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We aimed to develop and validate a Loop-mediated Isothermal Amplification (LAMP) assay to Sporothrix brasiliensis. LAMP reaction was developed using six primers designed based on calmodulin gene. In the LAMP reaction, we tested twenty isolates of S. brasiliensis from animals and humans, along with ten tissue samples extracted from the left footpad of mice that had been experimentally infected with S. brasiliensis. In addition, it included DNA samples from various other fungal species for specificity evaluation. All S. brasiliensis isolates yielded positive results in the LAMP, and the limit of DNA detection was 1 ng/µL. All murine samples were positive in the test while DNA from other fungal species were all negative, resulting in 100% of sensitivity and specificity of primers. LAMP diagnosis technique is a promising alternative to sporotrichosis diagnosis, in a simple and cost-effective way. Further studies are warranted to validate this technique using animal model samples obtained from both humans and animals.

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Sporotrichosis is a mycosis with zoonotic potential caused by species of Sporothrix. Once thought rare in northeastern Brazil, the disease has recently been spreading, leading to an emergency health issue. In this paper, we describe an outbreak of feline sporotrichosis in the Seventh Health District of Maceió-AL. We collected samples from 23 domiciled and non-domiciled felines without regard for age, breed, sex, and neutering state. Skin samples were analyzed cytologically under a light microscope and seeded onto Sabouraud dextrose agar at 25 °C for from 15 to 30 days. Fifteen of the twenty-three cats with suspected skin lesions were positive for Sporothrix spp. on either cytological or microbiological evaluation. Most of the infected cats were male, young adults, non-neutered, with free access to external areas, and living in environments with poor sanitation, a high population density, and an accumulation of garbage and organic matter. Three owners were bitten or scratched by infected cats and subsequently developed suspicious cutaneous lesions suggestive of sporotrichosis. The epidemiological features of feline sporotrichosis in the outbreaks of Maceió seemed to share similarities with the data obtained from outbreaks in current hyperendemic areas. Identifying geographical sites of infection and providing compulsory notification of the disease is essential for avoiding an epidemic in Alagoas.

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INTRODUCTION: Sporotrichosis is a subcutaneous mycosis caused by different species of thermodimorphic fungi belonging to the genus Sporothrix. These fungi may affect a wide range of warm-blooded animals, including humans, dogs and, mainly, cats. METHODS: In this article, we conducted a narrative literature review about the status of feline and cat-transmitted sporotrichosis in Latin American countries, with emphasis on Brazil. RESULTS: From a global perspective, Latin America is one of the main foci of human and feline sporotrichosis. In humans, the disease has been reported in several countries, with endemic transmission occurring in Brazil, Colombia, Costa Rica, Guatemala, Mexico and Uruguay. Sporothrix brasiliensis is the most common species identified in human cases in Brazil, whereas Sporothrix schenckii predominates in other countries. CONCLUSION: Domestic cats are the main source of infection in the zoonotic transmission cycle of sporotrichosis in Latin America. Cases of feline sporotrichosis have been reported in several countries, including Argentina, Brazil, Chile, Colombia, Mexico, Panama, Paraguay and Peru.

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We report Sporothrix brasiliensis infection in three cats from Santiago, Chile. Recently, S. brasiliensis was reported in cats from the southernmost region of Chile located 2,190 km from Santiago. Our findings emphasize the emergence of S. brasiliensis in the Chilean context, reflecting its rapid expansion across South America in recent years. Veterinarians should include S. brasiliensis in the differential diagnosis of skin conditions in cats.

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Sporotrichosis is a mycotic infection of the cutaneous and subcutaneous tissues caused by Sporothrix spp. that can also cause extracutaneous manifestations. This study aimed to characterize cutaneous and extracutaneous sporotrichosis lesions in cats. Over 1 year, 102 cats rescued by the Zoonoses Control Center of Belo Horizonte, Brazil, euthanized with clinical suspicion of feline sporotrichosis were evaluated. After euthanasia, the animals were evaluated by macroscopic, cytological, histopathological, and immunohistochemistry (IHC) examinations; fungal culture; and polymerase chain reaction (PCR). Sporothrix infection was identified by at least one diagnostic technique in all cats (n = 102) evaluated by postmortem examination, including 26/28 cases (93%) evaluated by IHC, 66/90 cases (73%) evaluated by cytology, 70/102 cases (68.6%) evaluated by histopathology, and 62/74 cases (84%) evaluated by fungal culture. Two cats had positive results only by fungal culture. Cytology and histopathology examinations were effective in diagnosing sporotrichosis, although IHC was needed to confirm the diagnosis in cats with low fungal loads. Sporothrix brasiliensis was confirmed by the sequencing of 3 samples. Skin lesions were characterized mainly by pyogranulomatous to granulomatous dermatitis (frequently with subcutaneous inflammation) with different intensities of Sporothrix spp. yeast. Extracutaneous findings associated with sporotrichosis included rhinitis or rhinosinusitis, lymphadenitis, pneumonia, meningitis, periorchitis, conjunctivitis, and glossitis. Extracutaneous infections were observed in 74/102 cases, and a possible association between the chronicity of the disease and the higher pathogenicity of this fungal species in cats requires further investigation.

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Background: Sporotrichosis caused by Sporothrix brasiliensis is a globally emerging infectious disease with limited therapeutic options. Thus, we aimed to evaluate the in vitro activity of amlodipine (AML) and lufenuron (LUF) alone and their interaction with itraconazole (ITZ), the first-choice drug against S. brasiliensis. Methods: Twenty clinical isolates of S. brasiliensis from two hyperendemic regions were tested through a microdilution assay to evaluate the minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of AML and LUF. Checkerboard assay was performed with 10 isolates for both drug interactions with ITZ. Results: AML showed inhibitory and fungicidal activity against all isolates included, with MIC values ranging from 32 to 256 µg/mL, and MFC from 64 to 256 µg/mL. However, none of the S. brasiliensis isolates were inhibited by the highest soluble concentration of LUF (MIC >64 µg/mL for all strains). Synergic interaction of AML and LUF with ITZ occurred in 50% and 40% of the isolates tested, without any antagonistic effects. Conclusion: Both repurposing drugs evaluated in our study showed a promising in vitro activity, especially in synergy with ITZ against S. brasiliensis, warranting future in vivo investigations regarding its activity.

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BACKGROUND: Sporotrichosis is a fungal infection that can affect both humans and animals, caused by a species of thermo-dimorphic fungi of the genus Sporothrix. This pathology can be acquired by subcutaneous traumatic inoculation through contact with contaminated plants, soil or decomposing organic matter, and/or by inhalation of conidia. The infection can progress to chronic skin infection, or it can even spread to blood vessels, lymph, muscles, bones, and other organs, such as the lungs and nervous system. Those disseminated types are usually associated with cellular immunodeficiency and infection by inhalation, which explains why people living with human immunodeficiency virus (PLHIV) get infected in such a manner. This virus changes the natural history of sporotrichosis, producing a greater fungal load. METHODS: The search was carried out in three databases: Pubmed, Scopus, and Scielo. Eligible articles were considered as those that described sporotrichosis in patients infected with HIV-AIDS, as well as case series. RESULTS: A total of 24 articles were selected, with a sum of 37 patients with sporotrichosis and HIV infection. Of these patients, 31 came from Brazil, two from the United States, one from South Africa, one from Bangladesh, and two from an unspecified region. Regarding epidemiology, a predominance of the male sex was found in 28 of the 37 cases (75.6%), while nine were female (24.3%). CONCLUSIONS: Sporotrichosis infection continues to present in a more severe and disseminated way among HIV-positive subjects with lower CD4+ counts.

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Introduction. For over a century, Sporothrix schenckii was considered the sole species responsible for sporotrichosis. In 2007, scientific community confirmed the disease could be caused by various Sporothrix species. These species differed in their virulence factors and their antifungal sensitivity. Objective. This study aims to characterize 42 Colombian clinical isolates of Sporothrix spp. phenotypically and genotypically. Materials and methods. Forty-two clinical isolates were characterized using phenotypic methods. It involved various culture media to determine their growth range at different temperatures and to assess the type and distribution of pigment and colony texture. Microscopic morphology was evaluated through microcultures, as well as the conidia diameter, type of sporulation, and morphology. Additionally, the assimilation of carbohydrates was selected as a physiological trait for species identification. Genotyping of 40 isolates was performed through partial amplification of the calmodulin gene, followed by sequence analysis. Results. Molecular studies enabled the identification of 32 isolates of S. schenckii and 8 isolates of S. globosa. The combination of phenotypic and genotypic methods eased these species characterizations and the recognition keys development based on parameters such as growth diameter at 25 and 30 °C, colony texture (membranous or velvety) on potato dextrose agar, and microscopic morphology with predominance of pigmented triangular, elongated oval globose, or subglobose conidia. Conclusions. Confirmation of the phenotypic characteristics and molecular analysis is crucial for identifying Sporothrix species and determining adequate treatment. This study represents the first phenotypical and genotypical characterization of clinical isolates of Sporothrix spp. reported in Colombia.

Introducción. Por más de un siglo se creyó que Sporothrix schenckii era la única especie responsable de la esporotricosis. Sin embargo, en el 2007, se consideró que podría ser causada por diferentes especies de Sporothrix, que difieren en sus factores de virulencia y su sensibilidad a los antifúngicos. Objetivo. Caracterizar fenotípica y genotípicamente 42 aislamientos clínicos colombianos de Sporothrix spp. Materiales y métodos. Se caracterizaron 42 aislamientos clínicos mediante métodos fenotípicos. Se usaron varios medios de cultivo para determinar el rango de crecimiento a diferentes temperaturas, el tipo y la distribución del pigmento, y la textura de las colonias. Se evaluó la morfología microscópica por microcultivos mediante la determinación del diámetro, el tipo de esporulación y la morfología de las conidias. La asimilación de carbohidratos se usó como una característica fisiológica para identificar las especies. La genotipificación de los 40 aislamientos se llevó a cabo mediante la amplificación parcial del gen que codifica para la calmodulina y se confirmó por secuenciación. Resultados. Mediante estudios moleculares, se identificaron 32 aislamientos de S. schenckii y ocho de S. globosa. La combinación de métodos fenotípicos y genotípicos permitió caracterizar las especies y construir claves para su reconocimiento, con base en parámetros como el diámetro de crecimiento a 25 y 30 °C, la textura de las colonias (membranosa, aterciopelada) en agar papa dextrosa y la morfología microscópica con predominio de conidias (triangulares pigmentadas, ovales globosas elongadas, subglobosas). Conclusiones. La caracterización fenotípica y los análisis moleculares son necesarios para identificar las especies de Sporothrix y, de esta forma, elegir el tratamiento indicado. Esta es la primera caracterización fenotípica y genotípica reportada de aislamientos clínicos colombianos de Sporothrix spp.

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Since 1998, the state of Rio de Janeiro, Brazil, has become a public health problem regarding sporotrichosis, a disease caused by Sporothrix spp. involving contact with infected cats. Efforts to isolate these species from environmental sources are not always successful. In our study, soil from residences situated in cities of Rio de Janeiro where cats with sporotrichosis live was collected and cultured an attempt to isolate Sporothrix spp. but it was not successful. However, other saprophytic fungal species were isolated from soil and identified and among them Purpureocillium lilacinum was the most frequent. From there, we decided to study the in vitro interaction of this species with S. brasiliensis, the principal agent that causes sporotrichosis in this state. The results showed that ten isolates of P. lilacinum inhibited the radial mycelial growth of S. brasiliensis with different percentage of inhibition. The interaction between them revealed the pattern described as overgrowth by antagonist. In conclusion, our data suggest that fungal species with very fast growth and capable of producing metabolites could hinder the growth of Sporothrix spp., it also opens the way for the identification of secondary metabolites with biological activity that could be tested against pathogenic fungi.

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Sporotrichosis is caused by fungi belonging to the genus Sporothrix, which saprophytically are found in plants and organic matter. However, cats are highly susceptible to contamination with fungal spores and, when they become sick, they can transmit it to other animals and to man. The objective of this study is to carry out a systematic review on the emergency, diagnosis, clinical symptoms, therapeutics, and control of zoonotic sporotrichosis. Published data covering the last 50 years using a combination of keywords were selected to answer the question: Why has the zoonotic sporotrichosis been a neglected disease up to now? A total of 135 studies were included in this review. The studies emphasize that in recent decades, Brazil has experienced an unprecedented zoonotic outbreak of sporotrichosis. Advances on the genus Sporothrix allowed one to associate thermotolerance, capacity for melanin synthesis, potential for adhesion to tissue macromolecules, ergosterol peroxide production, and expression of virulence proteins as tools for infection and invasion in S. brasiliensis, the main species involved, although cases with S. schenckii or S. lurei were also reported. Correct diagnosis, early treatment, basic educational measures that emphasize responsible ownership of animals and reproductive control programs for felines can contribute to the control of zoonosis.

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Perylene-based compounds, either naturally occurring or synthetic, have shown interesting biological activities. In this study, we report on the broad-spectrum antifungal properties of two lead amphiphilic perylene bisimides, compounds 4 and 5, which were synthesized from perylene-3,4,9,10-tetracarboxylic dianhydride by condensation with spermine and an ammonium salt formation. The antifungal activity was evaluated using a collection of fungal strains and clinical isolates from patients with onychomycosis or sporotrichosis. Both molecules displayed an interesting antifungal profile with MIC values in the range of 2-25 µM, being as active as several reference drugs, even more potent in some particular strains. The ammonium trifluoroacetate salt 5 showed the highest activity with a MIC value of 2.1 µM for all tested Candida spp., two Cryptococcus spp., two Fusarium spp., and one Neoscytalidium spp. strain. Therefore, these amphiphilic molecules with the perylene moiety and cationic ammonium side chains represent important structural features for the development of novel antifungals.

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Recently, sporotrichosis caused by Sporothrix brasiliensis transmitted during tattooing process was described in the epicenter of the Brazilian epidemic sporotrichosis. We report a similar case of this mycosis in a patient recently tattooed, but probably infected by a sick cat instead of via a contaminated procedure. Clinical cure was attained after two months of oral itraconazole. In the hyperendemic S. brasiliensis regions of sporotrichosis, health professionals must be aware of atypical transmissions of this fungus.

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BACKGROUND: Sporotrichosis is a disease not requiring jurisdictional notification and consequently is underreported in Brazil. Therefore, the epidemiological picture even in hyperendemic states is unknown. Thus, we evaluated the occurrence of sporotrichosis throughout the territory of the southern state of Brazil, Rio Grande do Sul (RS). METHODS: We update the epidemiological situation of sporotrichosis in the southern region of this state and describe the emergence of this disease in the Metropolitan region. We engaged professionals from RS enrolled in animal health care in answering a questionnaire regarding sporotrichosis. RESULTS: The occurrence of local cases of feline sporotrichosis was reported by 83% of the participants from 40 cities, distributed through the seven health districts of RS. Human sporotrichosis cases, transmitted by cats, were also reported by professionals from four regions of the state. The frequency of the disease in both the South and Metropolitan regions showed a marked increase in recent years. CONCLUSION: Feline and cat-transmitted human sporotrichosis is an underreported mycosis in RS, widely distributed in the territory of this state and increasing. Aggressive public health policies are urgently necessary to control the geographical expansion of this spreading mycosis.

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BACKGROUND: Sporotrichosis is a fungal implantation disease of subacute/ chronic course caused by species of the dimorphic fungus Sporothrix spp. This infection usually develops after traumatic inoculation of contaminated soil, plants or organic material contaminated by Sporothrix spp. conidia into skin or mucosa. The objective of this work is to contribute to the knowledge of sporotrichosis in Uruguay by providing a report of a series of cases diagnosed in a reference center. METHODS: We conducted a retrospective, observational, descriptive and cross-sectional study of cases of sporotrichosis diagnosed in the last 38 years. RESULTS: In the period analyzed, 157 cases of sporotrichosis were diagnosed, 152 of those corresponded to male patients. The most frequent clinical presentation was nodular lymphatic in 120 patients. In relation to epidemiological antecedents, 128 patients had been scratched by armadillos during hunting. CONCLUSIONS: Sporotrichosis in Uruguay is a sporadic disease with a clear seasonal pattern related to particular social practices, such as hunting armadillos. Related to this practice, the affectation is greater in males and in young adults.

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We describe the successful treatment of a series of 30 zoonotic sporotrichosis cases from southern Brazil. Sporothrix brasiliensis was the species genotypically identified in all 25 confirmed cases. Five other cases were classified as probable, without laboratory confirmation, but with clinical and epidemiological data of cat-transmitted sporotrichosis. Two isolates were sequenced by translation elongation factor-1 alpha (EF1α) loci in order to compare their sequences, and both of them showed distinct genotypes from S. brasiliensis strains from other Brazilian states. Itraconazole (ITZ) or potassium iodide (KI) were the first choice treatment in 28 and 2 cases, respectively. Microdilution assay showed a wild-type profile of S. brasiliensis isolates to ITZ. However, a lack of clinical response occurred in 42% of cases, especially those treated with ITZ 100 mg/day, and treatment needed modifications, by either increased doses or antifungal combinations. Clinical cure required a mean of 187 days of treatment, which was dependent on the clinical form of the disease and age of patients. Therapy, including dosages and durations, for cutaneous forms of sporotrichosis requires re-evaluation, since cases caused by S. brasiliensis may influence treatment efficacy.

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Sporothrix schenckii, Sporothrix brasiliensis, and Sporothrix globosa are the main causative agents of sporotrichosis, a human subcutaneous mycosis. Differences in virulence patterns are associated with each species but remain largely uncharacterized. The S. schenckii and S. brasiliensis cell wall composition and virulence are influenced by the culturing media, with little or no influence on S. globosa. By keeping constant the culturing media, we compared the cell wall composition of three S. schenckii and two S. brasiliensis strains, previously described as presenting different virulence levels on a murine model of infection. The cell wall composition of the five Sporothrix spp. strains correlated with the biochemical composition of the cell wall previously reported for the species. However, the rhamnose-to-ß-glucan ratio exhibits differences among strains, with an increase in cell wall rhamnose-to-ß-glucan ratio as their virulence increased. This relationship can be expressed mathematically, which could be an important tool for the determination of virulence in Sporothrix spp. Also, structural differences in rhamnomannan were found, with longer side chains present in strains with lower virulence reported for both species here studied, adding insight to the importance of this polysaccharide in the pathogenic process of these fungi.

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Sporotrichosis has become an important zoonosis in Brazil, and Sporothrix brasiliensis is the primary species transmitted by cats. Improvement of animal treatment will help control and limit the spread and geographic expansion of sporotrichosis. Accordingly, buparvaquone, an antiprotozoal hydroxynaphthoquinone agent marketed as Butalex, was evaluated in vitro and in vivo against feline-borne isolates of S. brasiliensis. Buparvaquone inhibited in vitro fungal growth at concentrations 4-fold lower than itraconazole (the first-choice antifungal used for sporotrichosis) and was 408 times more selective for S. brasiliensis than mammalian cells. Yeasts treated with a subinhibitory concentration of buparvaquone exhibited mitochondrial dysfunction, reactive oxygen species and neutral lipid accumulation, and impaired plasma membranes. Scanning electron microscopy images also revealed buparvaquone altered cell wall integrity and induced cell disruption. In vivo experiments in a Galleria mellonella model revealed that buparvaquone (single dose of 5 mg/kg of body weight) is more effective than itraconazole against infections with S. brasiliensis yeasts. Combined, our results indicate that buparvaquone has a great in vitro and in vivo antifungal activity against S. brasiliensis, revealing the potential application of this drug as an alternative treatment for feline sporotrichosis.

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Sporotrichosis is a mycotic disease caused by Sporothrix spp., whose zoonotic transmission by sick cats is the main infection route in Brazil. The aim of the current study is to report a human sporotrichosis outbreak caused by zoonotic transmission from a feline, with emphasis on the importance of making differential diagnosis and of using personal protective equipment. A hospital team member presented injury in the arm after having handled a cat that had been hospitalized for being hit by a car. The animal presented skin lacerations, myiasis, and full tibial fracture - there were no other signs of skin lesions. Clinical samples were collected from both the human and the suspected cat, for mycological culture; results have shown Sporothrix sp. growth. A search was conducted to identify other hospital team members who also had contact with the animal. Other six individuals also had suspected lesions in their arms, hands and ocular area; they were all subjected to sample collection. Mycological results have also confirmed Sporothrix spp.; sequencing analysis has shown that all seven humans were infected with Sporothrix brasiliensis. Since Southern Brazil is endemic of this disease, it is worth emphasizing the importance of taking into consideration zoonotic risks at the time to provide emergency care to stray animals, mainly felines, as well as of using Personal Protective Equipment while handling them - regardless of whether they present, or not, typical clinical symptoms or history of the disease, given the potential zoonotic risk posed by Sporothrix brasiliensis.

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Sporotrichosis is an endemic mycosis caused by the species of the Sporothrix genus, and it is considered one of the most frequent subcutaneous mycoses in Mexico. This mycosis has become a relevant fungal infection in the last two decades. Today, much is known of its epidemiology and distribution, and its taxonomy has undergone revisions. New clinical species have been identified and classified through molecular tools, and they now include Sporothrix schenckii sensu stricto, Sporothrix brasiliensis, Sporothrix globosa, and Sporothrix luriei. In this article, we present a systematic review of sporotrichosis in Mexico that analyzes its epidemiology, geographic distribution, and diagnosis. The results show that the most common clinical presentation of sporotrichosis in Mexico is the lymphocutaneous form, with a higher incidence in the 0-15 age range, mainly in males, and for which trauma with plants is the most frequent source of infection. In Mexico, the laboratory diagnosis of sporotrichosis is mainly carried out using conventional methods, but in recent years, several researchers have used molecular methods to identify the Sporothrix species. The treatment of choice depends mainly on the clinical form of the disease, the host's immunological status, and the species of Sporothrix involved. Despite the significance of this mycosis in Mexico, public information about sporotrichosis is scarce, and it is not considered reportable according to Mexico's epidemiological national system, the "Sistema Nacional de Vigilancia Epidemiológica." Due to the lack of data in Mexico regarding the epidemiology of this disease, we present a systematic review of sporotrichosis in Mexico, between 1914 and 2019, that analyzes its epidemiology, geographic distribution, and diagnosis.

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Sporotrichosis is a subcutaneous granulomatous disease caused by the fungus Sporothrix spp. In Brazil, S. brasiliensis is reported in regions of outbreaks and epidemics in the zoonotic form of the disease where cats play an important role in the transmission of the disease to humans. Therefore, it is important to assess how the presence of infected cats impacts the risk for sporotrichosis in humans. The objective of this study was to analyze the spatial association of sporotrichosis in cats and in humans from Belo Horizonte, a Brazilian city where an epidemics of sporotrichosis occurs since the first human case register in 2015, through an inhomogeneous Poisson process model. Feline and human cases of sporotrichosis recorded between January 2016 and June 2019 were georeferenced by address and spatial point patterns were generated. Feline case intensity and human demographic density were calculated using a kernel smoothed estimate. The distance to the nearest feline case was also compute. Model parameters were estimated by Maximum Likelihood Estimate. The model validation was performed by the evaluation of partial residual, leverage and influence measure. There were 343 cases of cats and 135 human cases of sporotrichosis. The average incidence of human sporotrichosis in the period was 1.343 per 100 thousand inhabitants, which is relatively low in relation to the population, but higher than that observed in other regions in zoonotic outbreak of the disease. The southern region of the municipality has a higher intensity of feline cases. According to the fitted model, the risk for human sporotrichosis is greater when at distances very close to a feline case, with a virtually stable effect for distances greater than 1 km. Regarding the intensity of feline cases there is a gradual increase in risk as the intensity of cases increases. From the leverage analysis it was observed that the model was particularly sensitive to the occurrence of human cases in the south and east regions, places with extreme values ​​of covariates. Poisson point process model seems to be a reasonable approach in spatial epidemiology when multiple sources of infection are involved, and there is a low incidence of the disease as long as it is reasonable to assume independence between cases. Interventions for disease prevention and control in humans are suggested to encompass disease control in cats and the search for feline cases, focused on diagnosis and control, close to reported human cases.

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