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The severity of lesions that develop in patients infected by Leishmania braziliensis is mainly associated with a highly cytotoxic and inflammatory cutaneous environment. Recently, we demonstrated that senescent T and NK cells play a role in the establishment and maintenance of this tissue inflammation. Here, we extended those findings using transcriptomic analyses that demonstrate a strong co-induction of senescence and pro-inflammatory gene signatures in cutaneous leishmaniasis (CL) lesions. The senescence-associated signature was characterized by marked expression of key genes such as ATM, Sestrin 2, p16, p21 and p38. The cell type identification from deconvolution of bulk sequencing data showed that the senescence signature was linked with CD8+ effector memory and TEMRA subsets and also senescent NK cells. A key observation was that the senescence markers in the skin lesions are age-independent of patients and were correlated with lesion size. Moreover, a striking expression of the senescence-associated secretory phenotype (SASP), pro-inflammatory cytokine and chemokines genes was found within lesions that were most strongly associated with the senescent CD8 TEMRA subset. Collectively, our results confirm that there is a senescence transcriptomic signature in CL lesions and supports the hypothesis that lesional senescent cells have a major role in mediating immunopathology of the disease.

Background Paracoccidioidomycosis (PCM) is an endemic disease in Latin America. In immunocompetent hosts, PCM occurs in two main clinical forms: acute and chronic. However, in HIV-infected patients PCM may show up simultaneous manifestations of acute and chronic forms. Case report We present the case of a patient diagnosed with HIV who had disseminated skin lesions and generalized lymphadenopathy, as well as respiratory and central nervous system involvement. The PCM diagnosis was confirmed by direct KOH examination, double immunodiffusion and the isolation of the fungus in samples of an abscess in the subcostal region. The isolate was identified as Paracoccidioides brasiliensis S1 by species-specific PCR using primers for protein-coding gene GP43 (exon 2) followed by PCR-RFLP of the alpha-tubulin gene. Conclusions There are few data in literature reporting species-specific molecular identification of Paracoccidioides in HIV/PCM patients. Therefore, this case report may contribute to improve the knowledge about this severe disease, its causative cryptic species, and its consequences to patients.

Background Sporotrichosis is a subcutaneous mycosis caused by Sporothrix species that affects humans and animals. Little information on the genetic diversity and population structure of the pathogen is available for Brazil, which is needed to design effective strategies to tackle the advance of sporotrichosis in endemic areas. Objectives We assessed the genetic diversity and mating-type distribution of Sporothrix isolates recovered from human and feline cases of sporotrichosis in Espirito Santo-Brazil to better understand the population structure, epidemiology, and diversification of this pathogen, as well as to explore the possible routes of transmission involved in the ongoing outbreaks. Methods In all, 75 Sporothrix isolates were identified with phenotypic characteristics. Then, fungal DNA extraction was performed, and the species-specific PCR technique was applied, using markers directed to the calmodulin gene. The mating-type idiomorph of species was identified by PCR using primers targeting the MAT1-1 and MAT1-2 loci. Results Among the 75 Sporothrix isolates, 76% were confirmed as S. brasiliensis and 24% as S. schenckii sensu stricto. S. brasiliensis was more prevalent in the metropolitan area and S. schenckii s. str. in the mountainous region of the state. In both species, the presence of the two sexual idiomorphs was detected, suggesting that they are heterothallic species. Conclusions Our data suggest that sporotrichosis takes on an epidemic-urban character involving S. brasiliensis. This species in Espirito Santo is likely to originate from Rio de Janeiro, as most isolates harbor the same MAT 1-2 locus. We confirm that S. brasiliensis has significantly broadened its area of occurrence, an essential feature of emerging pathogens.

Paracoccidioidomycosis (PCM) is an endemic systemic mycosis that is of great importance in Latin America. Its occurrence in solid organ transplantation (SOT) is rare, but with high mortality rate. In this report, we describe a case of PCM in a liver transplant recipient 19 months after transplantation. The patient presented with multiple skin abscesses, arthritis, osteolytic lesions, and pulmonary and adrenal involvement. Despite the presence of disseminated disease and the patient’s immunosuppressed condition, the patient responded well to prolonged antifungal treatment with no sequelae, thus suggesting that early diagnosis and correct treatment may lead to favorable outcomes in SOT recipients with PCM.

This article shows reports of the clinical–epidemiological characteristics and serological screening in patients assisted by a reference center for PCM care, University Hospital Cassiano Antonio Moraes, Federal University of Espirito Santo, Brazil. The patient’s sera with PCM were analyzed by DID test at the beginning and the end treatment. Clinical and demographic data were also collected to characterize the sample. One hundred patients with a suspected diagnosis of PCM were evaluated. Serology by DID test was used as a screen in all patients. The test was positive for 79 patients (72 for Paracoccidioides brasiliensis and 7 for Paracoccidioides lutzii). Serology was negative in 21 sera, although all of them were diagnosed PCM by histopathologic or direct exam. Serological follow-up was performed during the treatment of all patients. After treatment, 58(58%) had negative serology and 33(33%) low levels of antibodies (≤ 1:16). Our results indicate the importance of the DID test for the screening and monitoring of PCM and that the incidence of P. lutzii might be greater than expected in areas where it is not the predominant PCM species. Therefore, this article may contribute to improving the knowledge and clinical management about this disease.

Leishmaniasis is a tropical infectious disease caused by Leishmania spp. protozoa and is transmitted by insects from the Phlebotominae subfamily. It can manifest as cutaneous leishmaniasis, a painless ulcer that can develop into a more serious systemic affliction as the protozoa spreads lymphatically or hematogenously, depending on the host's immunity. In this case series, the authors present a rare form of genital mucocutaneous leishmaniasis, with consideration of epidemiologic characteristics, clinical presentation, differential diagnosis, and treatments offered.

Visceral leishmaniasis (VL) is an infectious disease caused by the protozoa Leishmania chagasi, whose main vector in South America is Lutzomyia longipalpis. The disease was diagnosed in the Brazilian state of Espirito Santo (ES) for the first time in 1968. Currently, this disease has been considered endemic in 10 municipalities. Furthermore, the presence of L. longipalpis has been detected in eight other municipalities where the transmission has not been reported thus far. In this study, we performed species distribution modeling (SDM) to identify new and most likely receptive areas for VL transmission in ES. The sandflies were both actively and passively collected in various rural area of ES between 1986 and 2017. The collection points were georeferenced using a global positioning system device. Climatic data were retrieved from the WorldClim database, whereas geographic data were obtained from the National Institute for Space Research and the Integrated System of Geospatial Bases of the State of Espirito Santo. The maximum entropy algorithm was used through the MIAmaxent R package to train and test the distribution models for L. longipalpis. The major contributor to model generation was rocky outcrops, followed by temperature seasonality. The SDM predicted the expansion of the L. longipalpis-prone area in the Doce River Valley and limited the probability of expanding outside its watershed. Once the areas predicted suitable for L. longipalpis occurrence are determined, we can avoid the inefficient use of public resources in conducting canine serological surveys where the vector insect does not occur.

Background: Paracoccidioidomycosis (PCM) is the most prevalent endemic systemic fungal infection in Latin America. In Brazil, it stands out as the eighth-highest cause of mortality among chronic or recurrent infections and has the highest mortality rate among systemic mycoses. Oral mucosal lesions may be the first visible physical manifestation of the disease. This study traced the epidemiological and clinical profiles of patients with oral lesions treated at the University Hospital Cassiano Antonio Moraes, Federal University of Espirito Santo. Methods: A retrospective cross-sectional analysis of patient medical records was performed. Results: Among the 161 patients identified with a confirmed diagnosis of PCM, 97 (60.24%) presented with oral lesions. The male:female ratio was 15:1, the mean age was 50.5 years, and the chronic form of paracoccidioidomycosis was predominant. Most of the patients had smoking habits and were rural workers. The most common oral lesions present in various anatomical sites were mulberry-like ulcers, more frequently observed in the gingiva, with regression within one to three months. Patients completed the treatment in one to two years (32.99%), and 47.42% of cases discontinued treatment. Conclusions: In addition to the characteristics of the oral lesions, information from the clinical profiles of patients with oral PCM is a central tool for dentists for early diagnosis. Earlier diagnosis may result in fewer consequences, especially respiratory ones that may cause an inability to work and poor quality of life. Keywords: Paracoccidioidomycosis, Oral lesions, Epidemiology, Clinical manifestations

Paracoccidioidomycosis is an endemic disease in Latin America that is rarely associated with immunosuppression and biological therapy. Herein, we report for the first time a case of pulmonary paracoccidioidomycosis reactivation after infliximab treatment. A 47-year-old man from Brazil received infliximab to treat psoriatic spondyloarthropathy and presented with cough, dyspnea, weight loss, and fever. Chest computed tomography revealed a pulmonary nodule and biopsy confirmed paracoccidioidomycosis. Treatment with sulfamethoxazole and trimethoprim was initiated for fungal infection and infliximab was reintroduced two months later. Considering his clinical improvement and favorable radiologic evolution, antifungal therapy was discontinued after 29 months.

In the south and southeast of Brazil, autochthonous malaria cases can be found near Atlantic Forest fragments. The transmission is not totally clarified; thus, the behaviour of the possible vectors in those regions must be observed. An entomological and natural infection study was performed on anophelines (Diptera: Culicidae) captured in the municipalities of the mountainous region of Espírito Santo state in 2004-2005. Similarly, between the years 2014 and 2015, 12 monthly collections were performed at the permanent trapping station of the study mentioned above (Valsugana Velha, Santa Teresa, ES). Light traps with CO2 (CO2-baited Center for Disease Control [CDC] traps) were set in open areas, at the edge of the forest (canopy and ground) and inside the forest (canopy and ground), whereas Shannon traps were set on the edge of the forest. A total of 1,414 anophelines were collected from 13 species. Anopheles (Kerteszia) cruzii Dyar and Knab remained the most captured species in the CO2-baited CDC traps set in the forest canopy and was also the vector with the highest prevalence of Plasmodium vivax infection according to molecular PCR techniques. Regarding mosquitoes of the subgenus Nyssorhynchus, P. vivax was found only in abdomens, weakening the hypothesis that this subgenus also plays a role in malaria transmission in this specific region.SponsorshipEspírito Santo Research Foundation (Fundação de Amparo à Pesquisa e Inovação do Espírito Santo – FAPES).

Paracoccidioidomycosis (PCM) is an endemic disease in the southeast region of Brazil, which includes the state of Espírito Santo (ES). This historic case series analyses 546 patients treated in this state from 1978 to 2012. Patients aged from 7 to 83 years, 509 males and 37 females, yielding a ratio of 13.7:1. Most of the patients (81.4%) originated from rural areas in ES, 71.0% being farmers. A higher concentration of cases was observed in municipalities located along the western range of the state. Sixty patients (11.0%) had an acute/subacute form of the disease, 485 (88.9%) had a chronic form, and one had a subclinical form. The most affected organs included the lungs, oropharyngeal mucosa, lymph nodes, skin, and larynx. The diagnosis was confirmed by histopathology in 252 (46.2%) cases, direct examination in 168 (30.7%), both exams in 111 (20.3%) and serology in 15 (2.8%). Tuberculosis, acquired immune deficiency syndrome, leishmaniasis, and intestinal parasites were the most frequently associated infectious diseases. From 328 patients followed up, total regression of the lesions was observed in 17.4%, partial regression in 77.4%, and no regression in 17 (5.2%) cases. Regarding the number of cases in this series, ES emerges as an important endemic area for PCM in Brazil.

Paracoccidioidomycosis (PCM) is a fungal disease caused by Paracoccidioides spp., which can cause a systemic granulomatous infection with tegumentary and visceral involvement. Sarcoid-like skin lesions are uncommon and can be misdiagnosed due to similarities with other granulomatous diseases. We report a case of a women presenting with erythematous infiltrated plaques on her face that was treated for leprosy and rosacea with no response and was later diagnosed with PCM, presenting positive serology for Paracoccidioides lutzii.

Cytotoxic activity mediated by CD8+ T cells is the main signature of the immunopathogenesis of cutaneous leishmaniasis (CL). Here, we performed a broad evaluation of natural killer (NK) cell phenotypic and functional features during cutaneous leishmaniasis. We demonstrate for the first time that CL patients present the accumulation of circulating NK cells with multiple features of replicative senescence including low proliferative capacity and shorter telomeres, elevated expression of CD57, KLRG1 but diminished CD27 stimulatory receptor expression. Moreover, they exhibited higher cytotoxic and inflammatory potential than agematched controls. The accumulation of circulating senescent NK cells (CD56dim CD57bright) correlated positively with skin lesion size in the same patients, suggesting that they, like circulating senescent CD8+ T cells, may contribute to the immunopathology of CL. However, this senescent population had lower cutaneous lymphocyte antigen expression and so had diminished skin-homing potential compared with total or senescent CD8+ T cells. This was confirmed in CL skin lesions where we found a predominance of CD8+ T cells (both senescent and non-senescent) that correlated with the severity of the disease. Although there was also a correlation between the proportions of senescent NK cells (CD56+ CD57+ ) in the skin and lesion size, this was less evident. Collectively our results demonstrate first-hand that senescent cytotoxic cells may mediate skin pathology during human cutaneous leishmaniasis. However, as senescent cytotoxic CD8+ T cells predominate in the skin lesions, they may have a greater role than NK cells in mediating the non-specific skin damage in CL.

BACKGROUND AND PURPOSE: Paracoccidioidomycosis is a fungal infection mainly caused by the thermodimorphic fungus Paracoccidioides. The purpose of our study was to demonstrate the neuroimaging findings from 24 patients with CNS paracoccidioidomycosis. MATERIALS AND METHODS: We performed a retrospective analysis focusing on the radiologic characteristics of CNS paracoccidioidomycosis. The 24 selected patients underwent MR imaging and/or CT, and the diagnosis was made by the presence of typical neuroimaging features, combined with fungus isolation, a serologic test, or the presence of disseminated disease. RESULTS: Headache was the most common neurologic symptom, while the pseudotumoral form was the most common pattern. The number of lesions ranged from 1 to 11, with most localized on the frontal lobe with >2-cm lesions. CT showed mainly hypoattenuating lesions, whereas MR imaging demonstrated mainly hyposignal lesions on T1WI and T2WI. Furthermore, ring enhancement was present in most patients. The “dual rim sign” on SWI occurred in 100% of our patients with lesions of >2 cm. CONCLUSIONS: The diagnosis of CNS paracoccidioidomycosis is difficult. Nevertheless, imaging examinations can play an important role in the diagnosis and evaluation of the disease.

International audience; The yellow fever virus (YFV) caused a severe outbreak in Brazil in 2016-2018 that rapidly spread across the Atlantic Forest in its most populated region without viral circulation for almost 80 years. A comprehensive entomological survey combining analysis of distribution, abundance and YFV natural infection in mosquitoes captured before and during the outbreak was conducted in 44 municipalities of five Brazilian states. In total, 17,662 mosquitoes of 89 species were collected. Before evidence of virus circulation, mosquitoes were tested negative but traditional vectors were alarmingly detected in 82% of municipalities, revealing high receptivity to sylvatic transmission. During the outbreak, five species were found positive in 42% of municipalities. Haemagogus janthinomys and Hg. leucocelaenus are considered the primary vectors due to their large distribution combined with high abundance and natural infection rates, concurring together for the rapid spread and severity of this outbreak. Aedes taeniorhynchus was found infected for the first time, but like Sabethes chloropterus and Aedes scapularis, it appears to have a potential local or secondary role because of their low abundance, distribution and infection rates. There was no evidence of YFV transmission by Aedes albopictus and Aedes aegypti, although the former was the most widespread species across affected municipalities, presenting an important overlap between the niches of the sylvatic vectors and the anthropic ones. The definition of receptive areas, expansion of vaccination in the most affected age group and exposed populations and the adoption of universal vaccination to the entire Brazilian population need to be urgently implemented.

This study discusses a historical patient series and is designed to describe clinical and epidemiological characteristics of human sporotrichosis in the state of Espirito Santo, Brazil. Data were derived from patients treated at the Infectious Diseases service of Cassiano Antonio Moraes University Hospital in Vitoria, the state capital, from July 1982 to June 2012. A total of 171 patients were diagnosed with sporotrichosis, mostly men (80.7%) with a median age of 33 years and 5 months. We can presume an approximate average incidence rate of 4.9 cases per 100,000 inhabitants during the studied period. All the patients were involved in occupational or leisure activities with direct contact with soil or plants. Most cases were recorded in the mountainous region of the state during the hot and rainy periods. The average time elapsed from lesion progression to diagnosis was 3 months, with the lymphocutaneous form being the most common (70.2%), followed by the fixed cutaneous form (28.6%). Diagnosis was confirmed in 93.6% of the cases by culturing Sporothrix spp. in Sabouraud dextrose agar, and from the clinical features in the remaining cases. Aspiration of cutaneous nodule secretions was the best method for the collection of clinical specimens for disease diagnosis. A 25% saturated solution of potassium iodide (SSKI) was provided to almost all patients (98.8%), with therapeutic success. In conclusion, in this retrospective study in the state of Espirito Santo, we found that sporotrichosis affects primarily the ≥ 10-year-old population, and the most common presentation is the lymphocutaneous form affecting the lower and upper limbs, and the infection appeared to be acquired predominantly through occupational activities. Treatment with SSKI was safe and effective.

Mucocutaneous leishmaniasis and paracoccidioidomycosis are infectious diseases with similar epidemiological and clinical aspects. Cases of both diseases may manifest similar lesions in the mucosa. Therefore, the determination of distinguishing characteristics for the purpose of differential diagnosis is critical for better management of the diseases. The present study evaluated factors that assist in the differentiation of mucosal lesions between these diseases. This cross-sectional study included data from medical records of 122 cases of mucocutaneous leishmaniasis and 83 cases of paracoccidioidomycosis attended at the university hospital Cassiano Antonio Moraes, located in Vitoria, Espirito Santo State, Brazil. Comparison between the diseases included the following variables: sex, age, time of disease evolution, location of the lesion and symptoms. Adults and males were affected by both diseases at higher rates. Lesions in the nasal region (95.1%; p-value = 0.000) and the pharynx (20.5%; p-value = 0.009) and nasal obstruction (34.4%; p-value = 0.000) were associated with leishmaniasis. Paracoccidioidomycosis was associated with lesions in the oral region (90.4%; p-value = 0.000), oral pain (16.9%; p-value = 0.000), and hoarseness (14.5%; p-value = 0.008). In leishmaniasis, lesions in oral regions were not associated with oral pain and were frequently located close to the nasal area. The manifestations cited above could improve the differential diagnosis of leishmaniasis and paracoccidioidomycosis, and thereby potentially aid in the choice of appropriate confirmatory diagnostic testing.

Paracoccidioidomycosis is an infectious disease characterized primarily by pulmonary involvement and potential dissemination to other organs, mainly mucosa and skin; however, it can affect any organ in the body. Although difficult to diagnose purely based on imaging, imaging is important for diagnosis, follow-up, and assessment of disease-related complications. We provide a comprehensive review of the most notable imaging findings of paracoccidioidomycosis.

A 59-year-old female patient, previously healthy, immunocompetent, presented left bulbar conjunctiva lesions and nodular-ulcerated lesions on the arms and cervical region, besides left cervical and retroauricular lymphadenopathy. She had previous contact with domestic cats that excoriated her face. The diagnosis was conclusive of disseminated sporotrichosis through clinical and epidemiological history and cultures of skin and ocular secretions. It evolved with good response to oral antifungal therapy.

BackgroundThe transmission of malaria in the extra-Amazonian regions of Brazil, although interrupted in the 1960s, has persisted to the present time in some areas of dense Atlantic Forest, with reports of cases characterized by particular transmission cycles and clinical presentations. Bromeliad-malaria, as it is named, is particularly frequent in the state of Espírito Santo, with Plasmodium vivax being the parasite commonly recognized as the etiologic agent of human infections. With regard to the spatial and temporal distances between cases reported in this region, the transmission cycle does not fit the traditional malaria cycle. The existence of a zoonosis, with infected simians participating in the epidemiology, is therefore hypothesized. In the present study, zoonotic transmission of bromeliad-malaria in Espírito Santo is investigated, based on the complete mitochondrial genome of DNA extracted from isolates of Plasmodium species which had infected humans, a simian from the genus Allouata, and Anopheles mosquitoes. Plasmodium vivax/simium was identified in the samples by both nested-PCR and real-time PCR. After amplification, the mitochondrial genome was completely sequenced and compared in a haplotype network, including all sequences of P. vivax/simium mitochondrial genomes sampled from humans and simians from all regions in Brazil.ResultsThe haplotype network demonstrates that humans and simians from the Atlantic Forest share the same haplotype, but some isolates from humans are not identical to the simian isolate. In addition, the plasmodial DNA extracted from mosquitoes revealed sequences different from those obtained from simians, but similar to two isolates from humans.ConclusionsThese findings reinforce the hypothesis that in the Atlantic Forest, and especially in the state with the highest frequency of bromeliad-malaria in Brazil, the same parasite species is shared by humans and simians, at least in part. The difference between the sequences of mosquitoes and simians raises two hypotheses: (1) two distinct transmission cycles for human malaria exist in the study area, one of them involving simians and the other exclusive to human hosts, or (2) there is only one transmission cycle involving humans and simians, but the identification of variations among simians was not possible due to a lack of other samples.

Knowledge of the fauna composition of anopheline mosquitoes, their ecological aspects and behavior, and influence of climatic variables on their population dynamics can help in understanding the transmission of Plasmodium parasites and thus develop more efficient strategies for the control of malaria. In the Central Atlantic Forest Biodiversity Corridor, southeastern Brazil, foci of introduced malaria have been reported among people returning from the Amazon region, north Brazil. Our objective was to evaluate and compare the anopheline fauna from a preserved environment and an adjacent peridomiciliary modified environment at the Central Atlantic Forest Biodiversity Corridor. We collected anopheline mosquitoes on a monthly basis from June 2004 to May 2006 from both these environments to understand the ecological aspects and their association with the occurrence of malaria. We captured 5,491 anopheline mosquitoes belonging to two subgenera and 11 species and studied the correlations between anopheline mosquito species and climatic variables. We considered Anopheles darlingi (Root) as the principal malaria vector and Anopheles albitarsis s. l. (Arribalzaga) as the secondary vector.

Amphotericin B deoxycholate is the main option for intravenous (i.v.) treatment of severe paracoccidioidomycosis (PCM). This is the first report of amphotericin B lipid complex (ABLC) in the treatment of PCM. Among 28 patients, cure was achieved in all patients (100%) using ABLC. Mean and median daily doses of ABLC were 3.39 mg/kg/day and 3.35 mg/kg/day, respectively. ABLC may be a choice in the treatment of severe forms of PCM or when i.v. options are required.

Cutaneous leishmaniasis (CL) is caused by a protozoan of the genus Leishmania and is transmitted by sand flies. The state of Espirito Santo (ES), an endemic area in southeast Brazil, has shown a considerably high prevalence in recent decades. Environmental niche modelling (ENM) is a useful tool for predicting potential disease risk. In this study, ENM was applied to sand fly species and CL cases in ES to identify the principal vector and risk areas of the disease. Sand flies were collected in 466 rural localities between 1997 and 2013 using active and passive capture. Insects were identified to the species level, and the localities were georeferenced. Twenty-one bioclimatic variables were selected from WorldClim. Maxent was used to construct models projecting the potential distribution for five Lutzomyia species and CL cases. ENMTools was used to overlap the species and the CL case models. The Kruskal–Wallis test was performed, adopting a 5% significance level. Approximately 250,000 specimens were captured, belonging to 43 species. The area under the curve (AUC) was considered acceptable for all models. The slope was considered relevant to the construction of the models for all the species identified. The overlay test identified Lutzomyia intermedia as the main vector of CL in southeast Brazil. ENM tools enable an analysis of the association among environmental variables, vector distributions and CL cases, which can be used to support epidemiologic and entomological vigilance actions to control the expansion of CL in vulnerable areas.

To assess the effect of the rapid removal of potentially infectious dogs on the prevalence and incidence of canine infections, a prospective study was undertaken in an area endemic for Leishmania infantum. We used serological testing based on the rapid DPP rK28 fusion protein chromatographic immunoassay for this dog screening-and-culling intervention trial. The outcome was evaluated by measuring seropositivity and sero-conversion/-reversion rates for canine infection. Our estimates indicated that concomitant detection and elimination of seropositive dogs with active disease may affect the numbers of canine infections and disease burden temporarily, although it is insufficient as a measure to interrupt the zoonotic L. infantum transmission. However, most of the asymptomatic, seropositive dogs continuously exhibit low levels of antibodies and/or reverted, remaining seronegative thereafter. In the process of waiting for an effective vaccine, one option for canine reservoir control may be to identify these possibly genetically resistant animals and promote their expansion in the population.

Introduction Sporotrichosis is a mycosis caused by the dimorphic fungus Sporothrix spp., and the primary mode of transmission is traumatic inoculation of fungus conidia from plants and other organic matter. Methods Sporotrichosis was established in a cat and in three members of one family from a rural area, by fungal growth in culture. Results The cutaneous lesions developed on the lower buttock and limbs with no report of any injury. Conclusion Although sporotrichosis should be considered for differential diagnosis in people working with soil and plants, cases of chronic lesions on skin of cats and in households is probably a new epidemiological approach in a given geographic region.

Canine visceral leishmaniasis (CVL) is the major source of human visceral leishmaniasis (VL) and is transmitted from dogs to sand flies to humans. To control the spread of this disease, early and accurate detection of infected dogs is critical but challenging. Here we demonstrate the potential of the Dual-Path Platform (DPP(®)) CVL rapid test for detecting K26/K39-reactive antibodies in sera from clinically symptomatic (n=60) and asymptomatic (n=60) Leishmania infantum-infected dogs. For the specificity evaluation, assays were performed using known negative diagnostic serum samples (n=59) and cross-reaction control sera (n=11) from animals born in a VL-free area of Brazil. The diagnostic kit displayed high specificity (96%) but low sensitivity (47%) in identifying parasite-positive dogs without signs of CVL. However, the test sensitivity was significantly higher (98%) in diseased cases, indicating that this convenient test may be useful to identify the most infectious dogs. Efforts should be pursued to obtain a more sensitive DPP-multiplexed test parameter (i.e. based on simultaneous yet separate antibody detection of carefully selected multiple antigens of diagnostic utility) for effective serodiagnosis of early-infected dogs, as this will likely allow more efficient canine removal regimens than those used in practice by public health services.

Free-living amoeba of the genus Acanthamoeba can eventually act as parasites, causing infections in humans. Some physiological characteristics of Acanthamoeba have been related to the grade of pathogenicity, allowing inferences about the pathogenic potential. The main goal of this study was to characterize isolates of Acanthamoeba obtained in Brazil and evaluate properties associated with their pathogenicity. A total of 39 isolates obtained from keratitis cases (n = 16) and environmental sources (n = 23) were classified into morphological groups and genotyped by sequencing the 18S rDNA fragments ASA.S1 and GTSA.B1. Samples were also tested regarding their thermo-tolerance, osmo-tolerance, and cytopathogenicity in MDCK cells. Isolates were identified and classified as follows: group I (T17, T18); group II (T1, T3, T4, T11); and group III (T5, T15), with the predominance of genotype T4 (22/39). Clinical isolates were genotyped as T3 (1/16), T4 (14/16) and T5 (1/16). The majority of isolates (38/39) were able to grow at 37 °C, but tolerance to 40 °C was more frequent among environmental samples. The tolerance to 1 M mannitol was infrequent (4/39), with three of these corresponding to clinical samples. The variable ability to cause cytopathic effects was observed among isolates of distinct genotypes and origins. This study identified, for the first time, T1, T15, and T18 in Brazil. It also indicated a weak association between the clinical origin of the isolates and tolerance to high temperatures, high osmolarity, and cytopathogenicity, demonstrating that some in vitro parameters do not necessarily reflect a higher propensity of Acanthamoeba to cause a disease.

In an endemic rural area of southeast Brazil, surveys confirmed that dogs serve as peridomestic reservoirs of Leishmania infantum. It is likely that the lack of efficient control is because presently used diagnostic tests miss positive dogs. Overall, 57% of the dogs had specific antibodies, but the canine infection was not uniformly fatal and many seropositive dogs remained asymptomatic or even spontaneously recovered. Furthermore, 42% of the human residents became leishmanin-positive reactors and 47% had positive serology at the initial survey, but our estimates also point at a high recovery rate among the infected population with time. The delayed-type hypersensitivity (DTH) reaction to Leishmania was a good indicator of resistance to infection in this particular epidemiologic situation. The lack of any significant differences in infection rates by gender or age indicate that all of the population was at an equal risk of infection and most people were infected in the peridomestic setting.

The diagnosis of visceral leishmaniasis remains difficult in rural areas where the disease is endemic, and serologic methods still need assessment, as they are not very sensitive for the detection of asymptomatic infectious dogs. Here we present data on the development of enzyme-linked immunosorbent assay (ELISA)-based methods for the detection of antibodies against recombinant leishmanial antigens (namely, the recombinant K26 [rK26] and rK39 antigens from Leishmania infantum and the rA2 protein from Leishmania donovani ) in comparison to ELISAs employing crude soluble antigen (CSA). The assays utilized sera from known negative controls ( n = 25) and clinically asymptomatic ( n = 50) and symptomatic ( n = 50) dogs with confirmed L. infantum infections. Additional studies were also done using sera from animals harboring other infections ( n = 14) for the evaluation of cross-reactivity. Our study indicated that rK26 and rK39 used in ELISAs provided very high sensitivities for the detection of symptomatic dogs (94% and 100%, respectively), followed by CSA (88%) and rA2 (70%). Conversely, rA2 was more sensitive for asymptomatic dogs (88%) than rK39 and rK26 (both 66%) and CSA (30%). Some cross-reactivity in sera from dogs with other infections ( Leishmania braziliensis and Leptospira interrogans ) was identified, but the rA2 protein provided the greatest specificity (98%). Data further indicate that all three recombinant proteins must be used in parallel to detect essentially all infected dogs. Efforts should be made to develop a cheap and reliable serologic test based on epitope selection from these diagnostic markers for the sensitive detection of L. infantum -infected dogs.

In Brazil, the main etiologic agent of Leishmaniasis that frequently presents with mucosal involvement belongs to the Viannia subgenus. The therapeutic conduct in this disease depends on the parasitological diagnosis, and classical methods are restricted in identifying the agent. In this paper we describe a polymerase chain reaction (PCR), which uses primers designed from mini-exons repetitive sequences. The PCR amplifies a 177bp fragment that can distinguish (Viannia) from (Leishmania) subgenus. This test could be a useful diagnostic tool.

Background: Because domestic dogs are reservoir hosts for visceral leishmaniasis (VL) in Brazil, one of the approaches used to reduce human disease incidence is to cull infected dogs. However, the results of controlled intervention trials based on serological screening of dogs and killing of seropositive animals are equivocal. A prophylactic vaccine to protect dogs from being infectious to the sand fly vector could be an effective strategy to provide sustained control. Here, we investigated whether a currently licensed commercial subunit rA2 protein-saponin vaccine (Leish-tec®) had an additional effect to dog culling on reducing the canine infectious populations. Methodology/principal findings: This prospective study was conducted in an L. infantum highly endemic area of southeast Brazil. At the onset of the intervention, all of the eligible dogs received through subcutaneous route a three-dose vaccine course at 21-day intervals and a booster on month 12. For the purpose of comparison, newly recruited healthy dogs were included as the exposed control group. To ascertain vaccine-induced protection, dogs were screened on clinical and serological criteria every 6 months for a 2-year follow-up period. Antibody-based tests and histopathological examination of post-mortem tissue specimens from euthanized animals were used as a marker of infection. The standardized vaccine regime, apart from being safe, was immunogenic as immunized animals responded with a pronounced production of anti-A2-specific IgG antibodies. It should be noted the mean seroconversion time for infection obtained among immunized exposed dogs (~ 18 months), which was twice as high as that for unvaccinated ones (~ 9 months). After two transmission cycles completed, the cumulative incidence of infection did differ significantly (P = 0.016) between the vaccinated (27%) and unvaccinated (42%) dogs. However, the expected efficacy for the vaccine in inducing clinical protection was not evident since 43% of vaccine recipients developed disease over time. Our estimates also indicated that immunoprophylaxis by Leish-tec® vaccine in addition to dog culling might not have an impact on bringing down the incidence of canine infection with L. infantum in areas of high transmission rates. Conclusions/significance: Leish-tec® as a prophylactic vaccine showed promise but needs to be further optimized to be effective in dogs under field conditions, and thereby positively impacts human incidence.

Introduction and objectives: Jorge Lobo’s disease is a rare chronic disease caused by the fungus Lacazia loboi, characterized by keloid lesions on the skin. The objective is to perform a review of the literature and report the first case of Jorge Lobo’s disease treated at the University Hospital Cassiano Antonio Moraes (HUCAM). Materials and methods: The study data were obtained retrospectively from the analysis and systematic review of the medical records of a patient with Jorge Lobo’s disease identified in HUCAM. Case Report: A male patient aged 37 years was referred to HUCAM in April 2010 presenting with keloid lesions and a previous diagnosis of lobomycosis by histopathology examination. On December 2011 a partial removal of the lesions with grafting was performed. Three months later, there was recurrence of lobomycosis on the grafted tissue. In June 2012 drug treatment with clofazimine associated with fluconazole was started. The lack of satisfactory clinical response after two months meant that the disease was considered beyond cure. Discussion and literature review: lobomycosis is an endemic disease in the brazilian Amazon and other humid tropical areas of Central and South America, and its transmission occurs by inoculation of the fungus through breaks in the skin. Diagnosis is made by clinical examination and histopathologiy or cytology The choice of therapy varies with the extent of lesions. However, none of the therapies described in the literature is fully effective. Conclusion: In this case, the two main therapeutic approaches failed, and the disease was considered incurable.

The involvement of different sand fly species in the transmission of American cutaneous leishmaniasis (ACL) at different altitudes was evaluated in the municipality of Afonso Cláudio in the State of Espírito Santo, Brazil, from November 1995 to February 1997. CDC light traps and Shannon traps baited with human volunteers were hung simultaneously at three altitudes of a river valley: (a) 650-750 m in an area of active ACL transmission; (b) 750-850 m in a transitional area; and (c) 850-950 m in an area where no ACL transmission occurred. A total of 13,363 specimens belonging to 28 species was collected. The five most abundant man-biting species were Lutzomyia intermedia, which constituted 24.3% of the total, Lu. migonei (22.3%), Lu. whitmani (15.4%), Lu. fischeri (14.9%) and Lu. monticola (5.8%). Analysis of the distribution of these species at the three altitudes provided evidence that Lu. fischeri and Lu. monticola were not involved in ACL transmission, whereas Lu. migonei and Lu. whitmani might act as secondary vectors of Leishmania (Viannia) braziliensis and Lu. intermedia was probably the principal vector of the parasite. These results reinforce those published in the existing literature, which indicate that Lu. intermedia is the main vector of Le. (V.) braziliensis in Southeast Brazil, while Lu. migonei and Lu. whitmani are of secondary importance.