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Abstract: Visceral leishmaniasis (VL) is one of the most important tropical diseases worldwide. Although chemotherapy has been widely used to treat this disease, problems related to the development of parasite resistance and side effects associated with the compounds used have been noted. Hence, alternative approaches for VL control are desirable. Some methods, such as vector control and culling of infected dogs, are insufficiently effective, with the latter not ethically recommended. The development of vaccines to prevent VL is a feasible and desirable measure for disease control; for example, some vaccines designed to protect dogs against VL have recently been brought to market. These vaccines are based on the combination of parasite fractions or recombinant proteins with adjuvants that are able to induce cellular immune responses; however, their partial efficacy and the absence of a vaccine to protect against human leishmaniasis underline the need for characterization of new vaccine candidates. This review presents recent advances in control measures for VL based on vaccine development, describing extensively studied antigens, as well as new antigenic proteins recently identified using immuno-proteomic techniques.

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Abstract: Our colleagues at the Sociedade Brasileira de Medicina Tropical have been informed of the demise of Professor Pierre Ambroise-Thomas (1937-2014). However, considering that the tribute we paid to him in 2015 - at the 20th anniversary of the Seminário Laveran & Deane sobre Malária - is equally true today, it is worth sharing it with the readers of the RSBMT, in recognition of his many virtues. Pierre Ambroise-Thomas (MD in 1963 and DSc in 1969) was Honorary Professor of Parasitology and Tropical Medicine at the Faculté de Médecine de Grenoble (France), Honorary President of the Académie Nationale de Médecine, member of the Académie Nationale de Pharmacie and Officier dans l'Ordre de La Légion d'Honneur. In addition to his important contributions to tropical medicine and parasitology, working in France during his long and productive career (50 years and 300 publications), Ambroise-Thomas became an admirer and supporter of Brazilian activities related to research, teaching and information in Tropical Medicine.

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Abstract: INTRODUCTION: The morbidity and mortality profile in a given region reflects its quality of life and provides tools for improving public health policies in that region. METHODS A cross-sectional epidemiological study was performed using secondary mortality data collected from the Monte Negro municipality of the Brazilian Western Amazon from 2000 to 2011. These data were compared with data from similar municipalities in other Brazilian macro-regions. Data were obtained through the Departamento de Informática do Sistema Único de Saúde (DATASUS) information system. RESULTS The number of deaths reported over the study period was 606. The most common cause of death was external causes of morbidity and mortality [International Classification of Diseases (ICD)-10 chapter 20], followed by diseases of the circulatory system (ICD-10 chapter 9). Among the causes of death according to age group, infectious and parasitic diseases were the most common for 2- to 9-year-old children; external causes of disease were the most prevalent for 10- to 59-year-old people; and circulatory diseases prevailed in individuals over 60 years of age. Eleven percent of deaths were due to unknown causes. CONCLUSIONS These results point to a fragility in the public policies for prevention and awareness of this problem. Infectious and parasitic diseases contribute only 4.5% of deaths, but had the third highest Disability-Adjusted Life Year score (1,190 days). Improving support to the Estratégia Saúde da Família (Family Health Strategy) program and implementing a death verification service would significantly aid in reducing the occurrence of non-transmissible chronic diseases and clarifying unknown causes of death.

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Abstract: INTRODUCTION Information regarding the cost of implementing insecticide-treated curtains (ITCs) is scarce. Therefore, we evaluated the ITC implementation cost, in addition to the costs of intensive conventional routine activities of the Aedes control program in the city of Guantanamo, Cuba. METHODS A cost-analysis study was conducted from the perspective of the Aedes control program, nested in an ITC effectiveness trial, during 2009-2010. Data for this study were obtained from bookkeeping records and activity registers of the Provincial Aedes Control Programme Unit and the account records of the ITC trial. RESULTS The annual cost of the routine Aedes control program activities was US$16.80 per household (p.h). Among 3,015 households, 6,714 ITCs were distributed. The total average cost per ITC distributed was US$3.42, and 74.3% of this cost was attributed to the cost of purchasing the ITCs. The annualized costs p.h. of ITC implementation was US$3.80. The additional annualized cost for deploying ITCs represented 19% and 48.4% of the total cost of the routine Aedes control and adult-stage Aedes control programs, respectively. The trial did not lead to further reductions in the already relatively low Aedes infestation levels. CONCLUSIONS At current curtain prices, ITC deployment can hardly be considered an efficient option in Guantanamo and other comparable environments.

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Abstract: INTRODUCTION: Behavioral fever is a response to infections with microorganisms observed in some poikilothermic animals. Rhodnius prolixus is involved in the transmission of two parasites: Trypanosoma cruzi (pathogenic for humans and transmitted in feces) and Trypanosoma rangeli (non-pathogenic for humans, pathogenic for Rhodnius and transmitted by the bite of an infected individual). Only T. rangeli is found in the hemolymph of Rhodnius as it travels to the salivary glands. METHODS: To study vector-parasite interactions, we evaluated possible behavioral fever responses of R. prolixus to intracoelomic inoculation with T. cruzi or T. rangeli. Temperature preferences of fifth-instar nymphs of R. prolixus were evaluated after inoculation with T. rangeli KP1(+), KP1(-), T. cruzi I, or the Trypanosome culture medium. Four different fixed temperatures (25, 30, 35, and 40°C) in two simultaneous experiments (enclosed and free-moving insects) were evaluated. Free-moving insects were marked daily according to their temperature preferences on each of the 15 days after inoculation. Numbers of insects in each temperature shelter and daily mortality were compared with those enclosed shelters of different temperatures. RESULTS: Rhodnius prolixus inoculated with both strains of T. rangeli and with the trypanosome culture medium showed preferences for the lowest temperatures (25°C). However, R. prolixus inoculated with T. cruzi I showed significant preferences for temperatures around 35°C. CONCLUSIONS: This is the first known investigation to demonstrate a behavioral fever response in R. prolixus injected intracoelomically with T. cruzi I.

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Abstract: INTRODUCTION: Members of the Acinetobacter genus are key pathogens that cause healthcare-associated infections, and they tend to spread and develop new antibiotic resistance mechanisms. Oxacillinases are primarily responsible for resistance to carbapenem antibiotics. Higher rates of carbapenem hydrolysis might be ascribed to insertion sequences, such as the ISAba1 sequence, near bla OXA genes. The present study examined the occurrence of the genetic elements bla OXA and ISAba1 and their relationship with susceptibility to carbapenems in clinical isolates of the Acinetobacter calcoaceticus-Acinetobacter baumannii complex. METHODS: Isolates identified over 6 consecutive years in a general hospital in Joinville, Southern Brazil, were evaluated. The investigation of 5 families of genes encoding oxacillinases and the ISAba1 sequence location relative to bla OXA genes was conducted using polymerase chain reaction. RESULTS: All isolates presented the bla OXA-51-like gene (n = 78), and 91% tested positive for the bla OXA-23-like gene (n = 71). The presence of ISAba1 was exclusively detected in isolates carrying the bla OXA-23-like gene. All isolates in which ISAba1 was found upstream of the bla OXA-23-like gene (n = 69) showed resistance to carbapenems, whereas the only isolate in which ISAba1 was not located near the bla OXA-23-like gene was susceptible to carbapenems. The ISAba1 sequence position of another bla OXA-23-like-positive isolate was inconclusive. The isolates exclusively carrying the bla OXA-51-like gene (n = 7) showed susceptibility to carbapenems. CONCLUSIONS: The presence of the ISAba1 sequence upstream of the bla OXA-23-like gene was strongly associated with carbapenem resistance in isolates of the A. calcoaceticus-A. baumannii complex in the hospital center studied.

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Abstract: INTRODUCTION: Extended-spectrum beta-lactamases (ESBLs) are bacterial enzymes capable of hydrolyzing beta-lactams. The aim of this study was to describe the prevalence of TEM- and SHV-type ESBL-producing Klebsiella pneumoniae strains in Zahedan, Southeast Iran. METHODS: A total of 170 non-repetitive K. pneumoniae strains were collected from patients referred to three teaching hospitals of Zahedan. Antibiotic susceptibility testing was determined for 17 antibiotics using the Kirby-Bauer disc diffusion method. The frequency of ESBL-producing strains was calculated, and minimum inhibitory concentrations of ESBL-producing strains were determined for cefotaxime, ceftazidime, ceftriaxone, and cefpodoxime. The presence of bla TEM and bla SHV genes was tested in all ESBL-producing strains using polymerase chain reaction and DNA sequencing. RESULTS: Among the 170 K. pneumoniae clinical isolates, 55 (32.4%) were ESBL producers; 92.7% (n=51) and 72.7% (n=40) of the isolates carried the bla SHV and bla TEM genes, respectively, and 67.3% (n=37) carried both genes. The sequencing results showed that all bla TEM types were bla TEM-1, except for two isolates that were bla TEM-104. The bla SHV types were bla SHV-1, bla SHV-11, bla SHV-12, bla SHV-99, bla SHV-108, and bla SHV-110. CONCLUSIONS: The percentage of bla TEM and bla SHV among ESBL-producing K. pneumoniae isolates from Zahedan is relatively high, indicating the need for further surveillance and consideration in antibiotic use. To the best of our knowledge, this is the first report of TEM-104-, SHV-99-, SHV-108-, and SHV-110-type ESBLs among clinical isolates of K. pneumoniae from Iran, and TEM-1, SHV-1, SHV-11, and SHV-12 appear to be the dominant ESBLs in this region.

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Abstract: INTRODUCTION: In the Brazilian Amazon, malaria infections are primarily caused by Plasmodium vivax. The only drug that kills the hypnozoite form of P. vivax is primaquine, thereby preventing relapse. However, treating glucose-6-phosphate dehydrogenase (G6PD)-deficient individuals with primaquine can lead to severe hemolysis. G6PD deficiency (G6PDd) affects approximately 400 million people worldwide, most of whom live in malaria-endemic areas. Therefore, clinicians need tools that can easily and reliably identify individuals with G6PDd. This study estimated the accuracy of the Carestart(tm) G6PD rapid test (Access Bio) in the diagnosis of G6PDd in male participants with and without P. vivax acute malaria. METHODS: Male participants were recruited in Manaus. Malaria diagnosis was determined by thick blood smear. G6PD quantitative analysis was performed spectro photometrically at a wave length of 340nm. The Carestart(tm) G6PD test was performed using venous blood. Genotyping was performed for individuals whose samples had an enzyme activity less than 70% of the normal value. RESULTS: Six hundred and seventy-four male participants were included in this study, of whom 320 had a diagnosis of P. vivax malaria. In individuals with enzyme activity lower than 30% (n=13), the sensitivity, specificity, positive predictive value, and negative predictive value of the Carestart(tm) G6PD test were as follows: 61.5% (95%CI: 35.5%-82.3%), 98.3% (95%CI: 97.0%-99.1%), 42.1% (95%CI: 23.1%-63.7%), and 99.2% (95%CI: 98.2%-82.3%), 98.3% (95%CI: 97.0%-99.1%), 42.1% (95%CI: 23.1%-63.7%), and 99.2% (95%CI: 98.2%-99.7%), respectively. Increases in sensitivity were observed when increasing the cut-off value. CONCLUSIONS: Despite low sensitivity, Carestart(tm) G6PD remains a good alternative for rapid diagnosis of G6PDd in malaria-endemic regions.

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Abstract: INTRODUCTION Recent studies have shown a high incidence and prevalence of latent tuberculosis infection (LTBI) in indigenous populations around the World. We aimed to estimate the prevalence and annual risk of infection (ARI) as well as to identify factors associated with LTBI in an indigenous population from the Brazilian Amazon. METHODS We conducted a cross-sectional study in 2011. We performed tuberculin skin tests (TSTs), smears and cultures of sputum samples, and chest radiographs for individuals who reported cough for two or more weeks. Associations between LTBI (TST ≥5mm) and socio-demographic, clinical, and epidemiological characteristics were investigated using Poisson regression with robust variance. Prevalence ratio (PR) was used as the measure of association. RESULTS We examined 263 individuals. The prevalence of LTBI was 40.3%, and the ARI was 2.4%. Age ≥15 years [PR=5.5; 95% confidence interval (CI): 3.5-8.6], contact with tuberculosis (TB) patients (PR=3.8; 95% CI: 1.2-11.9), previous TB history (PR=1.4; 95% CI: 1.2-1.7), and presence of Bacillus Calmette-Guérin (BCG) scar (PR=1.9, 95% CI: 1.2-2.9) were associated with LTBI. CONCLUSIONS Although some adults may have been infected years prior, the high prevalence of infection and its strong association with age ≥15 years, history of TB, and recent contact with TB patients suggest that the TB transmission risk is high in the study area.

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Abstract: INTRODUCTION There is an increasing burden of multidrug resistance. As a result, deciphering the mechanisms of action of natural compounds with antifungal activity has gained considerable prominence. We aimed to elucidate the probable mechanism of action of citronellal, a monoterpenoid found in the essential oil extracted from Cymbopogon plants, against Candida albicans. METHODS Drug susceptibility was measured by broth microdilution and spot assays. Ergosterol levels were estimated using the alcoholic potassium hydroxide method and H+ extrusion was assessed by monitoring the glucose-induced acidification of the external medium. Virulence traits were studied by hyphal morphogenesis and biofilm formation, along with fungal cell adherence to polystyrene surface and human oral epithelial cells. RESULTS Citronellal showed anticandidal activity against C. albicans and non-albicans species of Candida at a minimum inhibitory concentration of 1 mg/ml. Citronellal interfered with membrane homeostasis, which is the major target of known antifungal drugs, by increasing the hypersensitivity of the fungi to membrane-perturbing agents, reducing ergosterol levels, and diminishing glucose-induced H+ extrusion. In addition, oxidative and genotoxic stresses were induced via an increased production of reactive oxygen species. Furthermore, citronellal inhibited the virulent attributes of yeast-to-hypha transition and biofilm formation. It also reduced cell adherence to polystyrene surface and the human oral epithelial cells. CONCLUSIONS This is the first study to propose the cell membrane, morphogenetic switching, biofilm formation, and cell adherence of Candida albicans as potential targets for the anticandidal activity of citronellal. However, clinical investigations on the therapeutic applications of citronellal are required.

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Abstract: INTRODUCTION: Rhinosporidiosis is a chronic infection of the mucous membrane and is caused by Rhinosporidium seeberi, an aquatic mesomycetozoan. The mode of infection is probably transepithelial penetration. The large number of rivers and lakes and the strong presence of riparian populations in the State of Maranhão are strong predisposing factors for rhinosporidiosis. METHODS: A 5-year retrospective study was conducted in a tertiary medical center situated in Maranhão, Northeast Brazil. Twenty-five Maranhense patients diagnosed with rhinosporidiosis were analyzed. RESULTS: Most of the patients were children, adolescents and young adults (age range: 7-24 years, mean age: 14 years). The majority of the participants were male (84%), brown (76%), and students (92%). All lesions involved the entire nasal cavity and presented with a vascular polypoid mass. All patients were treated by surgical excision of the lesions. CONCLUSIONS: Rhinosporidiosis affects younger age groups, especially students from the countryside and the outskirts of urban areas. This study will aid and guide physicians in diagnosing and treating this infection in endemic areas.

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Abstract: INTRODUCTION: Due to its frequency and morbidity, such as that caused by scorpions have achieved public health importance in certain regions of the world. The present exploratory ecological study aimed to characterize the epidemiological profile and spatial distribution of scorpion stings in Campina Grande, State of Paraíba in Northeastern Brazil. METHODS: Geographical information system techniques were used to record the scorpion stings, and Google Earth software, Track Maker, and ArcGIS 10 Esri were used as geocoding databases. The Moran test was used to evaluate spatial correlation, and the Pearson chi-square test was used to analyze associations between scorpion stings and socioeconomic variables. RESULTS: The study evaluated 1,466 scorpion stings. Envenomations were more frequent among women (n = 908, 61.9%), and most patients were aged 13-28 years (n = 428, 29.2%). The Southern region of the city had the largest number of registered cases (n = 548, 37.4%), followed by the Western region (n = 510, 34.8%). CONCLUSIONS: Spatial analysis of scorpionism revealed an irregular occurrence in Campina Grande. Further, no association was observed between the socioeconomic factors analyzed and the geographic location of the scorpion envenomations. Detection of spatial areas with an increased risk of scorpionism can help prioritize adoption of preventive measures in these regions to reduce the associated incidence and morbidity.

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Abstract: INTRODUCTION: Little information is available on the evolution of influenza A (H1N1)pdm09-related mortality in Brazil. METHODS: During the period 2009-2014, official epidemiological surveillance and mortality data were used to ascertain influenza-related deaths in the Santa Catarina State in Brazil. RESULTS: The highest overall mortality rate (2.3 per 100,000) was observed in 2009. Pregnant women constituted the highest risk group (mortality rate 18.0 per 100,000). Following vaccination of high-risk groups, the rate reduced sharply but peaked again in 2012. It subsequently decreased, albeit more gradually. CONCLUSIONS: Influenza A (H1N1)pdm09 mortality demonstrated a cyclic pattern, with two peaks followed by a gradual decrease.

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Abstract: INTRODUCTION: In this study, we evaluated hepatitis C virus (HCV) and human immunodeficiency virus (HIV) - platelet interactions in vitro as well as human platelets antigen (HPA) polymorphisms. METHODS: Platelets were obtained from 100 healthy HPA-genotyped volunteer donors and incubated with HIV or HCV. The viral load after in vitro exposure was detected. RESULTS: The viral load in the platelets after exposure to the virus was higher in the HIV exposure than in the HCV exposure. CONCLUSIONS: HIV-platelet ligation could be more efficient than HCV-platelet interaction. Further, the HPA-1b allele seems to influence the interaction of platelets with HCV.

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Abstract: INTRODUCTION: This study aimed to investigate Flavivirus infection in birds captured in green areas of São Paulo. METHODS: Oropharyngeal swabs, cloacal swabs, and blood samples from 170 birds captured in two green areas in São Paulo, Brazil were subjected to real time-polymerase chain reaction (RT-qPCR) analysis of Flavivirus specific NS5 gene fragment. RESULTS: All samples were negative for the presence of viral ribonucleic acid (RNA). CONCLUSIONS: Despite the negative results, Flavivirus surveillance must be performed regularly due to favorable ecological conditions for virus circulation and transmission among birds in these areas and their close proximity to humans.

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Abstract: INTRODUCTION: We report the behavioral changes in use of oviposition sites in Aedes aegypti in Fortaleza, Brazil. METHODS: We evaluated the relationship between different types of breeding sites and changes in use from 2001 to 2012. RESULTS: More than 40% of the infested breeding sites were used to store water. We observed a three-fold reduction in the infestation of water tanks (p = 0.038) and more than nine-fold in tires (p = 0.035). The proportion of infested plant pots increased six-fold (p < 0.001). CONCLUSIONS: Infested breeding sites changed over time from domestic water tanks to small-volume breeding sites.

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Abstract: INTRODUCTION: Triatomines are targeted for the control of Chagas disease in endemic areas of Brazil. METHODS: Data regarding triatomines captured during 2001-2008 in 34 municipalities in the Northeast of Minas Gerais were analyzed. RESULTS: In total, 11,187 triatomines from eight species were captured, mostly in henhouses and bedrooms. Trypanosomes were found in 203 samples. Main species were Triatoma sordida, Panstrongylus megistus, and Triatoma vitticeps. The number of P. megistus adults was positively correlated with temperature and precipitation, whereas the number of nymphs followed the inverse pattern. CONCLUSIONS: Occurrence of triatomines in domiciles indicates the need for sustained entomological surveillance.

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Abstract: INTRODUCTION: Mass production of mosquitoes under laboratory conditions allows implementing methods to control vector mosquitoes. Colony development depends on mosquito size and weight. Body size can be estimated from its correlation with wing size, whereas weight is more difficult to determine. Our goal was to test whether wing size can predict the weight. METHODS: We compared dry weight and wing centroid size of Culex quinquefasciatus reared at different temperatures and four diets. RESULTS: Weight and wing size were strongly correlated. The diets did not influence wing size. CONCLUSIONS: Wing centroid size is a good predictor of Cx. quinquefasciatus body weight.

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Abstract: INTRODUCTION Toxoplasmosis during pregnancy can be severe; thus, it is essential to diagnose the disease via serological tests. METHODS An enzyme-linked immunosorbent assay (ELISA) was used to investigate anti-Toxoplasma gondii immunoglobulin A (IgA), M (IgM) and G (IgG) antibodies in 62 high-risk pregnant women. RESULTS Forty-three (69.4%) women were positive for IgA, 31 (50%) for IgG, and 57 (91.9%) for IgM; 4 (6,5%) were positive for IgA but negative for IgM; 10 (16.1%) were negative for IgA and IgM but positive for IgG. CONCLUSIONS Testing for these antibodies can help diagnose infection in pregnant women, thereby contributing to clinical management.

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Abstract: INTRODUCTION: The monthly incidence of snakebites from 2007 to 2014 in the State of São Paulo, Brazil, was assessed. METHODS: A statistical model based on the discrete double Poisson distribution was proposed, including pairs of sine and cosine functions of time to account for seasonality and autoregressive terms. RESULTS: The analysis indicated a slight increase in the incidence of snakebites. The inclusion of two pairs of trigonometric functions seemed to be relevant in the model adjustment, given the seasonal pattern of the data. CONCLUSIONS: The snakebites occurred predominantly during the warm season, from November to April.

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Abstract: Because the majority of colubrid species are considered harmless to human beings, colubrid snakebites are rarely reported. However, the venom of Rhabdophis, which is part of the Colubridae family, is procoagulant and leads to severe coagulopathy. Here, we present a case of disseminated intravascular coagulation with enhanced fibrinolysis following a Rhabdophis bite. Although coagulopathy can be treated effectively with the specific Rhabdophis antivenom, this antivenom is not widely available in Indonesia. We also found transient hypertension secondary to the colubrid venom, an unusual finding.

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Abstract: We report herein a case of thoracic infection due to Nocardia nova following lung re-transplantation performed for emphysema related to alpha-1-antitrypsin deficiency. The infection extended from the lung into the pleural space, thoracic wall, and mediastinum, presenting as pericarditis and empyema necessitatis. Nocardia nova was identified by 16S ribosomal deoxyribonucleic acid (rDNA) sequencing and phylogenetic analysis. According to a literature search of PubMed, LILACS and MEDLINE databases, we describe herein the first case of empyema necessitatis caused by N. nova species in a transplanted patient.

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Abstract Sporotrichosis is the most common subcutaneous mycosis in South America and its association with zoonotic transmission remains a relevant public health problem in Rio de Janeiro, Brazil. The disease most commonly presents as subacute or chronic cutaneous lesions, although dissemination to various organs and systems occurs in rare cases, mainly in immunosuppressed individuals. This report describes a case of sporotrichosis with severe bone and subcutaneous damage in an immunocompetent patient who did not exhibit the characteristic skin lesions of sporotrichosis, including ulcers, nodules, and lymphangitis.

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Abstract: Injuries caused by fish are common in marine and freshwater environments. Catfish of the Ariidae and Pimelodidae families cause about 80% of those injuries. One of the complications of injuries caused by fish is the retention of fragments of the stinger in the wounds. Here we report five cases (of a total of 127 injuries caused by catfish in the Brazilian coast) in which the retained fragments were detected by radiological examination. Retained fragments should be considered in patients stung by catfish. A simple X-ray is sufficient to detect fragments of stingers in the wounds.