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1.
Mycopathologia ; 189(5): 73, 2024 Aug 03.
Article in English | MEDLINE | ID: mdl-39096410

ABSTRACT

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.


Subject(s)
DNA Primers , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques , Sensitivity and Specificity , Sporothrix , Sporotrichosis , Sporothrix/genetics , Sporothrix/isolation & purification , Sporothrix/classification , Sporotrichosis/diagnosis , Sporotrichosis/microbiology , Sporotrichosis/veterinary , Animals , Nucleic Acid Amplification Techniques/methods , Molecular Diagnostic Techniques/methods , Mice , Humans , DNA Primers/genetics , Disease Models, Animal , Calmodulin/genetics
2.
BMC Infect Dis ; 24(1): 824, 2024 Aug 14.
Article in English | MEDLINE | ID: mdl-39143511

ABSTRACT

BACKGROUND: Sporotrichosis is a chronic granulomatous infection of the skin and subcutaneous tissue that can affect any organ through lymphatic spread. The prevalence of sporotrichosis infections is increasing and its treatment is challenging as there are no unified and standard diagnostic techniques or antifungal medications. Controlling further spread requires a rapid diagnosis. Assessment of clinical symptoms, histological analysis, serological testing, and pathogen culture are all necessary for the diagnosis of sporotrichosis. However, these procedures are unable to identify the species. The development of safe, reliable, and species-specific diagnostic techniques is essential. OBJECTIVE: To establish and evaluate a new quantitative real-time PCR assay for the rapid diagnosis of sporotrichosis and to identify relevant species. METHODS: Polymorphisms in calmodulin (CAL) gene sequences and the internal transcribed spacer (ITS) were used in a quantitative real-time PCR assay to identify S. globosa, S. schenckii, and non-target species. RESULTS: The quantitative real-time PCR assay had 100% sensitivity and specificity. The limit of detection was 6 fg/µl. Thirty-four clinical specimens were verified to be infected with S. globosa with a 100% positive detection rate. CONCLUSIONS: The quantitative PCR technique developed in this study is a quick, accurate, and targeted method of identifying S. globosa based on polymorphisms in CAL sequences and ITS. It can be used for a prompt clinical diagnosis to identify S. globosa in clinical specimens from patients with sporotrichosis.


Subject(s)
Calmodulin , Real-Time Polymerase Chain Reaction , Sensitivity and Specificity , Sporothrix , Sporotrichosis , Sporotrichosis/diagnosis , Sporotrichosis/microbiology , Sporothrix/genetics , Sporothrix/isolation & purification , Humans , Real-Time Polymerase Chain Reaction/methods , Calmodulin/genetics , Asia , DNA, Fungal/genetics , Molecular Diagnostic Techniques/methods , Rapid Diagnostic Tests
3.
J Med Microbiol ; 73(8)2024 Aug.
Article in English | MEDLINE | ID: mdl-39145374

ABSTRACT

Introduction. Sporotrichosis is a subcutaneous infection caused by dimorphic Sporothrix species embedded in the clinical clade. Fungi have virulence factors, such as biofilm and melanin production, which contribute to their survival and are related to the increase in the number of cases of therapeutic failure, making it necessary to search for new options.Gap statement. Proton pump inhibitors (PPIs) have already been shown to inhibit the growth and melanogenesis of other fungi.Aim. Therefore, this study aimed to evaluate the effect of the PPIs omeprazole (OMP), rabeprazole (RBP), esomeprazole, pantoprazole and lansoprazole on the susceptibility and melanogenesis of Sporothrix species, and their interactions with itraconazole, terbinafine and amphotericin B.Methodology. The antifungal activity of PPIs was evaluated using the microdilution method, and the combination of PPIs with itraconazole, terbinafine and amphotericin B was assessed using the checkerboard method. The assessment of melanogenesis inhibition was assessed using grey scale.Results. The OMP and RBP showed significant MIC results ranging from 32 to 256 µg ml-1 and 32 to 128 µg ml-1, respectively. Biofilms were sensitive, with a significant reduction (P<0.05) in metabolic activity of 52% for OMP and 50% for RBP at a concentration of 512 µg ml-1 and of biomass by 53% for OMP and 51% for RBP at concentrations of 512 µg ml-1. As for the inhibition of melanogenesis, only OMP showed inhibition, with a 54% reduction.Conclusion. It concludes that the PPIs OMP and RBP have antifungal activity in vitro against planktonic cells and biofilms of Sporothrix species and that, in addition, OMP can inhibit the melanization process in Sporothrix species.


Subject(s)
Amphotericin B , Antifungal Agents , Melanogenesis , Proton Pump Inhibitors , Sporothrix , Sporotrichosis , Humans , Amphotericin B/pharmacology , Amphotericin B/therapeutic use , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Biofilms/drug effects , Biofilms/growth & development , Itraconazole/pharmacology , Melanins/biosynthesis , Melanins/metabolism , Melanogenesis/drug effects , Microbial Sensitivity Tests , Proton Pump Inhibitors/pharmacology , Proton Pump Inhibitors/therapeutic use , Sporothrix/drug effects , Sporothrix/metabolism , Sporotrichosis/drug therapy , Sporotrichosis/microbiology , Terbinafine/pharmacology
4.
PLoS Negl Trop Dis ; 18(7): e0012328, 2024 Jul.
Article in English | MEDLINE | ID: mdl-39038043

ABSTRACT

BACKGROUND: Sporotrichosis is the most common subcutaneous mycosis caused by Sporothrix spp. Traditionally, it is transmitted through injuries involving plant debris. However, over the past few decades, there has been an epidemic increase in human cases resulting from contact with infected animals, particularly cats, in various regions of Brazil. In this report, we report a notable increase in both human and animal cases within the Brazilian Amazon state. METHODOLOGY/PRINCIPAL FINDINGS: An ecological study was conducted by analyzing official records of human and animal sporotrichosis diagnosed in the state of Amazon from 2020 to 2023. Data including patient demographics, clinical manifestations, mycological examination results, and species identification through PCR confirmation were evaluated. During this period, a total of 950 human cases and 2,823 animal cases of sporotrichosis were reported at an exponential rate, since no human cases were registered in 2020. The spatial and temporal dispersion of human sporotrichosis followed that of animal cases, moving from downtown areas to the periphery. Contact with infected animals was reported in 77.7% of cases, with cats being the most commonly implicated (73.5%). Only 66.7% of individuals underwent mycological examination. Among the positive cultures for Sporothrix spp., 65.4% were identified as S. brasiliensis. All patients were treated with systemic antifungals. CONCLUSIONS/SIGNIFICANCE: This study highlights a rising incidence of sporotrichosis among animals and humans in the Brazilian Amazon region over the past four years, with S. brasiliensis being the predominant agent. Collaborative efforts involving healthcare professionals, veterinarians, and public health authorities are crucial to implement effective control measures, educate populations at risk, and promote responsible guidance for pet guardians. These measures are essential to mitigate the burden of epidemic sporotrichosis in Brazil.


Subject(s)
Disease Outbreaks , Sporothrix , Sporotrichosis , Zoonoses , Sporotrichosis/epidemiology , Sporotrichosis/microbiology , Sporotrichosis/veterinary , Brazil/epidemiology , Animals , Humans , Zoonoses/epidemiology , Zoonoses/microbiology , Zoonoses/transmission , Cats , Sporothrix/isolation & purification , Sporothrix/genetics , Male , Female , Adult , Public Health , Middle Aged , Adolescent , Young Adult , Aged , Child
6.
Med Mycol ; 62(8)2024 Aug 02.
Article in English | MEDLINE | ID: mdl-39043446

ABSTRACT

Zoonotic sporotrichosis caused by Sporothrix brasiliensis is an emerging mycosis in Latin America. One of the problems to quickly treat infected animals and break the transmission chain is associated with the time-consuming gold-standard diagnosis method (culture). We aimed to evaluate a species-specific polymerase chain reaction (PCR) for the diagnosis of sporotrichosis caused by S. brasiliensis using non-invasive samples. We performed a retrospective cross-sectional study using samples collected with swabs from humans and cats with clinical suspicion of sporotrichosis. Deoxyribonucleic acid (DNA) was extracted using a commercial kit, and a species-specific PCR for S. brasiliensis detection was performed. One hundred ten samples were included. PCR showed a good concordance with culture (86% of agreement) for human and cat samples (Kappa coefficient = 0.722, and 0.727, respectively). In conclusion, our data shows that this adapted PCR using non-invasive samples can be applied to sporotrichosis diagnosis, being a good alternative mainly in regions with a lack of mycologists to identify the fungus in culture, contributing to the control of this emergent zoonosis.


We aimed to evaluate a molecular method for diagnosing sporotrichosis caused by Sporothrix brasiliensis in humans and cats. We observed that the technique is in good agreement with the classic method and is a good alternative for assisting in the diagnosis and consequent control of this zoonosis.


Subject(s)
Cat Diseases , Polymerase Chain Reaction , Sporothrix , Sporotrichosis , Sporotrichosis/diagnosis , Sporotrichosis/microbiology , Sporotrichosis/veterinary , Cats , Sporothrix/genetics , Sporothrix/isolation & purification , Sporothrix/classification , Humans , Animals , Polymerase Chain Reaction/methods , Cat Diseases/diagnosis , Cat Diseases/microbiology , Retrospective Studies , Cross-Sectional Studies , Zoonoses/diagnosis , Zoonoses/microbiology , DNA, Fungal/genetics , Molecular Diagnostic Techniques/methods , Sensitivity and Specificity
7.
Prev Vet Med ; 230: 106275, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39043072

ABSTRACT

The research addresses zoonotic sporotrichosis in Brazil, particularly caused by Sporothrix brasiliensis, highlighting its epidemiological severity. Transmission occurs through contact with sick animals, especially felines, and diagnosis in humans is challenging due to the low fungal load in the lesions. The study analyzed data from Information System for Notifiable Diseases (SINAN) and Zoonosis Surveillance Unit (UVZ) from January 2017 to March 2023, carried out in Contagem, Minas Gerais. Geospatial tools and statistical analysis revealed a significant increase in cases, peaking in 2021 for felines and 2022 for humans. The geospatial analysis highlighted areas of higher incidence, suggesting a correlation between human and feline populations. The research contributes to the understanding of sporotrichosis in Contagem, emphasizing the importance of integrated approaches for surveillance and control strategies, aiming to mitigate impacts on the local community.


Subject(s)
Cat Diseases , Sporotrichosis , Zoonoses , Cats , Animals , Sporotrichosis/veterinary , Sporotrichosis/epidemiology , Sporotrichosis/microbiology , Cat Diseases/epidemiology , Cat Diseases/microbiology , Brazil/epidemiology , Zoonoses/epidemiology , Humans , Incidence , Spatio-Temporal Analysis , Sporothrix
8.
Future Microbiol ; 19(12): 1097-1106, 2024 Aug 12.
Article in English | MEDLINE | ID: mdl-39056139

ABSTRACT

Sporotrichosis, a mycosis resulting from cutaneous or subcutaneous infection with the dimorphic fungus Sporothrix, has been reported in China, particularly in the northeast region. In this review, we conducted a thorough examination of the recent advancements in sporotrichosis in China, encompassing aspects such as etiology, epidemiology, pathogenesis, clinical manifestations, diagnosis and treatment strategies. Within the Chinese context, fixed cutaneous sporotrichosis represents the prevailing clinical manifestation. Fungal culture stands as the gold standard for diagnosing sporotrichosis, while polymerase chain reaction techniques can enhance both the specificity and sensitivity of diagnosis. Besides conventional systemic antifungal agents, alternative modalities such as Chinese herbal medicines, photodynamic therapy and laser therapy show potential efficacy against sporotrichosis.


Sporotrichosis is a fungal infection on the skin. It is caused by a group of fungi called Sporothrix. Common symptoms are lesions on the skin, inflammatory papules, nodules and ulcers. These fungi live in the natural environment and cause infection by entering the body through a wound in the skin. China has a high prevalence of sporotrichosis, with northeast China the most seriously affected region. This review looks at the state of sporotrichosis in China.


Subject(s)
Antifungal Agents , Sporothrix , Sporotrichosis , Sporotrichosis/epidemiology , Sporotrichosis/microbiology , Sporotrichosis/drug therapy , Sporotrichosis/diagnosis , China/epidemiology , Humans , Sporothrix/isolation & purification , Sporothrix/genetics , Sporothrix/drug effects , Antifungal Agents/therapeutic use , Polymerase Chain Reaction , Photochemotherapy
9.
Mycopathologia ; 189(4): 66, 2024 Jul 14.
Article in English | MEDLINE | ID: mdl-39003373

ABSTRACT

Cat-transmitted sporotrichosis is caused by the emerging fungal pathogen Sporothrix brasiliensis and constitutes a significant public health issue that affects people living in resource-poor urban centers in Brazil. The lack of knowledge about transmission dynamics makes it difficult to propose public health policies to contain the advance of sporotrichosis. We describe the recent emergence of 1,176 cases of sporotrichosis in cats (2016 to 2021) in the metropolitan region of Recife, Brazil, leading to significant zoonotic transmission and an overwhelming occurrence of S. brasiliensis as the etiological agent. Most cases were from cats in the cities of Olinda (408/1,176; 34.70%), Jaboatão dos Guararapes (332/1,176; 28.23%), and Recife (237/1,176; 20.15%). Molecular typing using amplified fragment length polymorphism (EcoRI-GA/MseI-AG) revealed low polymorphic information content (PIC = 0.2499) and heterozygosity (H = 0.2928), typical of an outbreak scenario. Dendrogram and multivariate cluster analysis revealed that isolates from Pernambuco are closely related to Rio de Janeiro isolates. We report a substantial occurrence of MAT1-2 idiomorphs in the metropolitan region of Recife (0:60 ratio; χ2 = 60.000, P < 0.0001). The limited population differentiation and genetic diversity of the isolates from Pernambuco suggest a recent introduction, possibly via a founder effect, from the parental population in Rio de Janeiro. Our findings emphasize the critical importance of molecular surveillance of S. brasiliensis for outbreak response. A comprehensive one-health strategy is mandatory to control the spread of cat-transmitted sporotrichosis driven by S. brasiliensis, encompassing sanitary barriers, quick diagnosis, and treatment.


Subject(s)
Cat Diseases , Sporothrix , Sporotrichosis , Sporotrichosis/transmission , Sporotrichosis/microbiology , Sporotrichosis/veterinary , Sporotrichosis/epidemiology , Cats , Brazil/epidemiology , Sporothrix/genetics , Sporothrix/isolation & purification , Sporothrix/classification , Animals , Cat Diseases/microbiology , Cat Diseases/transmission , Cat Diseases/epidemiology , Molecular Typing , Zoonoses/transmission , Zoonoses/microbiology , Amplified Fragment Length Polymorphism Analysis , Communicable Diseases, Emerging/transmission , Communicable Diseases, Emerging/microbiology , Communicable Diseases, Emerging/epidemiology , Genotype , Phylogeny
10.
Acta Trop ; 257: 107297, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38914413

ABSTRACT

A 26-year-old woman presented an eyelid lesion, after being scratched by a cat that had a similar skin lesion. It evolved into a cervical lymph node enlargement. With a hypothesis of Parinaud´s oculoglandular syndrome (POS) due to cat scratch disease (CSD), doxycycline was prescribed. After two weeks of treatment without improvement, a biopsy and blood sample were obtained. Itraconazole was prescribed and the skin lesion improved, but not the lymph node enlargement. A Sporothrix schenckii complex was isolated from the skin sample. Also, a specie-specific polymerase chain reaction detected Bartonella henselae DNA in her blood sample. Azithromycin was included to treat the bacterial infection, whereupon the lymph node also receded successfully. Sporotrichosis and CSD are zoonoses that can be transmitted to humans by traumatic inoculation due to scratches or bites from cats. Both can evolve with POS. Patients who present skin lesions and/or POS after being wounded by a cat should be investigated for both diseases.


Subject(s)
Anti-Bacterial Agents , Bartonella henselae , Cat-Scratch Disease , Coinfection , Sporothrix , Sporotrichosis , Female , Bartonella henselae/isolation & purification , Bartonella henselae/genetics , Humans , Adult , Sporothrix/isolation & purification , Sporothrix/genetics , Sporotrichosis/microbiology , Sporotrichosis/drug therapy , Sporotrichosis/diagnosis , Coinfection/microbiology , Animals , Cats , Anti-Bacterial Agents/therapeutic use , Cat-Scratch Disease/microbiology , Cat-Scratch Disease/drug therapy , Cat-Scratch Disease/complications , Cat-Scratch Disease/diagnosis , Azithromycin/therapeutic use , Itraconazole/therapeutic use , Doxycycline/therapeutic use , Skin/microbiology , Skin/pathology
11.
Mycopathologia ; 189(4): 53, 2024 Jun 12.
Article in English | MEDLINE | ID: mdl-38864961

ABSTRACT

Sporotrichosis is a globally distributed subcutaneous mycosis caused by dimorphic Sporothrix species commonly found in soil, mosses, and decaying plant matter. The lymphocutaneous manifestation, historically associated with occupational activities and sapronotic transmission, has recently been observed to also occur through animal contact, particularly notable in Brazil. We describe a rare case of lymphocutaneous sporotrichosis with simultaneous pulmonary complications resulting from the scratching of a southern three-banded armadillo, Tolypeutes matacus, primarily inhabiting the arid forests of South America's central region. Speciation using multiplex quantitative polymerase chain reaction (qPCR) established the etiological agent as S. schenckii s. str., while amplified fragment length polymorphism (AFLP) analysis unveiled a novel genotype circulating in the Midwest of Brazil. The patient received treatment with itraconazole (200 mg/day) for two months, leading to substantial clinical improvement of cutaneous and pulmonary symptoms. This case highlights the critical role of animal-mediated transmission in sporotrichosis epidemiology, particularly within regions with diverse armadillo species. The unusual epidemiology and genetic characteristics of this case emphasize the need for enhanced awareness and diagnostic vigilance in atypical sporotrichosis presentations.


Subject(s)
Antifungal Agents , Armadillos , Itraconazole , Sporothrix , Sporotrichosis , Animals , Humans , Male , Amplified Fragment Length Polymorphism Analysis , Antifungal Agents/therapeutic use , Armadillos/microbiology , Brazil , Genotype , Itraconazole/therapeutic use , Multiplex Polymerase Chain Reaction , Sporothrix/genetics , Sporothrix/isolation & purification , Sporothrix/classification , Sporotrichosis/microbiology , Sporotrichosis/diagnosis , Sporotrichosis/drug therapy , Sporotrichosis/transmission , Treatment Outcome , Middle Aged
12.
Front Cell Infect Microbiol ; 14: 1382508, 2024.
Article in English | MEDLINE | ID: mdl-38828263

ABSTRACT

Introduction: Sporotrichosis is a subcutaneous mycosis caused by fungi of the genus Sporothrix sp. Phenotypic and genotypic differences have been associated with their geographic distribution, virulence, or clinical manifestation of sporotrichosis. In the past decade, the interest in identifying species of the Sporothrix sp. has been increasing, due to its epidemiological importance and, in consequence, is important to know how to preserve them for future studies, in culture collection. Aims: The purposes of this study were to analyze the global distribution of environmental isolates and/or causal agents of sporotrichosis identified by polyphasic taxonomy, with mandatory use of molecular identification, and to evaluate the percentages and distribution of isolates stored in culture collections. Methods: A systematic review of articles on animal and human sporotrichosis and/or environmental isolation of the fungus, from 2007 to 2023, was done. Results: Our results demonstrated that, S. globosa, S. schenckii, and S. brasiliensis were the most identified species. With respect to the deposit and maintenance of species, we observed that only 17% of the strains of Sporothrix sp. isolated in the world are preserved in a culture collection. Conclusions: This systematic review confirmed a difficulty in obtaining the frequency of Sporothrix species stored in culture collection and insufficient data on the molecular identification mainly of animal sporotrichosis and isolation of Sporothrix sp. in environmental samples.


Subject(s)
Sporothrix , Sporotrichosis , Sporothrix/classification , Sporothrix/isolation & purification , Sporothrix/genetics , Sporotrichosis/microbiology , Animals , Humans , Environmental Microbiology , Preservation, Biological/methods
13.
Curr Microbiol ; 81(7): 175, 2024 May 16.
Article in English | MEDLINE | ID: mdl-38755455

ABSTRACT

Vertebrate animal models are essential in research; however, efforts need to be made to decrease animal suffering as much as possible. It could be useful to determine humane endpoints that could serve as surrogates for a fatal outcome. We address this issue with respect to infectious diseases. We propose a humane endpoint for studies of Sporothrix brasiliensis infection. BALB/c mice were inoculated subcutaneously in the footpad. To define a humane endpoint, we considered two groups: animals who died during the experiment, and those euthanized at the end of the experiment. The groups were compared for colony-forming units (CFU) in internal organs, clinical condition, and body weight. Thirteen (22%) animals died before the end of the experiment due to the progression of local infection to severe and disseminated sporotrichosis. Analyzing data of the groups, we propose the following future criteria for euthanasia as a humane endpoint: neurological impairment OR necrosis of the footpad OR loss of ≥ 20% body weight AND moderate to severe dehydration. In view of the current global epidemiological impact of zoonotic sporotrichosis caused by S. brasiliensis, our data could facilitate the utility of models used to study the disease, particularly therapy.


Subject(s)
Disease Models, Animal , Mice, Inbred BALB C , Sporothrix , Sporotrichosis , Animals , Sporotrichosis/microbiology , Mice , Body Weight
14.
Emerg Microbes Infect ; 13(1): 2358073, 2024 Dec.
Article in English | MEDLINE | ID: mdl-38764403

ABSTRACT

Mycobacterium chelonae and Sporothrix globosa, both of which are opportunistic pathogens, have been proved to be possible multidrug resistant. However, are all recurring symptoms in chronic infections related to decreasing susceptibility? Here we report a case of sporotrichosis secondary to M. chelonae infection. In addition, we find that the blackish-red spots under the dermoscopic view can be employed as a signal for the early identification and regression of subcutaneous fungal infection.


Subject(s)
Mycobacterium Infections, Nontuberculous , Mycobacterium chelonae , Sporothrix , Sporotrichosis , Sporothrix/isolation & purification , Sporothrix/genetics , Sporothrix/drug effects , Sporotrichosis/microbiology , Humans , Mycobacterium chelonae/isolation & purification , Mycobacterium Infections, Nontuberculous/microbiology , Male , Coinfection/microbiology
15.
Emerg Infect Dis ; 30(5): 1053-1055, 2024 May.
Article in English | MEDLINE | ID: mdl-38666748

ABSTRACT

We describe a feline sporotrichosis cluster and zoonotic transmission between one of the affected cats and a technician at a veterinary clinic in Kansas, USA. Increased awareness of sporotrichosis and the potential for zoonotic transmission could help veterinary professionals manage feline cases and take precautions to prevent human acquisition.


Subject(s)
Cat Diseases , Sporotrichosis , Zoonoses , Animals , Cats , Female , Humans , Animal Technicians , Cat Diseases/microbiology , Cat Diseases/epidemiology , Cat Diseases/transmission , Kansas/epidemiology , Sporothrix/isolation & purification , Sporothrix/genetics , Sporotrichosis/veterinary , Sporotrichosis/transmission , Sporotrichosis/epidemiology , Sporotrichosis/microbiology , Zoonoses/epidemiology , Zoonoses/microbiology , Zoonoses/transmission
16.
Mycoses ; 67(4): e13724, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38584320

ABSTRACT

OBJECTIVE: This study aims to assess the clinical characteristics of sporotrichosis in low-endemic areas of China, including the prevalence geography, genotypic traits of patients, clinical manifestations, and strain virulence and drug sensitivities. The objective is to improve the currently used clinical management strategies for sporotrichosis. METHODS: Retrospective data were collected from patients diagnosed with sporotrichosis through fungal culture identification. The isolates from purified cultures underwent identification using CAL (Calmodulin) gene sequencing. Virulence of each strain was assessed using a Galleria mellonella (G. mellonella) larvae infection model. In vitro susceptibility testing against commonly used clinical antifungal agents for sporotrichosis was conducted following CLSI criteria. RESULTS: In our low-endemic region for sporotrichosis, the majority of cases (23) were observed in middle-aged and elderly women with a history of trauma, with a higher incidence during winter and spring. All clinical isolates were identified as Sporothrix globosa (S. globosa). The G. mellonella larvae infection model indicated independent and dose-dependent virulence among strains, with varying toxicity levels demonstrated by the degree of melanization of the G. mellonella. Surprisingly, lymphocutaneous types caused by S. globosa exhibited lower in vitro virulence but were more common in affected skin. In addition, all S.globosa strains displayed high resistances to fluconazole, while remaining highly susceptible to terbinafine, itraconazole and amphotericin B. CONCLUSION: Given the predominance of elderly women engaged in agricultural labour in our region, which is a low-epidemic areas, they should be considered as crucial targets for sporotrichosis monitoring. S. globosa appears to be the sole causative agent locally. However, varying degrees of melanization in larvae were observed among these isolates, indicating a divergence in their virulence. Itraconazole, terbinafine and amphotericin B remain viable first-line antifungal options for treating S.globosa infection.


Subject(s)
Sporothrix , Sporotrichosis , Aged , Middle Aged , Humans , Female , Itraconazole/pharmacology , Itraconazole/therapeutic use , Sporotrichosis/microbiology , Amphotericin B/pharmacology , Amphotericin B/therapeutic use , Terbinafine/therapeutic use , Retrospective Studies , Microbial Sensitivity Tests , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Sporothrix/genetics , China/epidemiology
17.
Braz J Microbiol ; 55(2): 1359-1368, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38466550

ABSTRACT

The drugs available to treat sporotrichosis, an important yet neglected fungal infection, are limited. Some Sporothrix spp. strains present reduced susceptibility to these antifungals. Furthermore, some patients may not be indicated to use these drugs, while others may not respond to the therapy. The anthelmintic drug niclosamide is fungicidal against the Sporothrix brasiliensis type strain. This study aimed to evaluate whether niclosamide also has antifungal activity against Sporothrix globosa, Sporothrix schenckii and other S. brasiliensis strains with distinct genotypes and antifungal susceptibility status. Minimal inhibitory and fungicidal concentrations (MIC and MFC, respectively) were determined using the microdilution method according to the CLSI protocol. The checkerboard method was employed to evaluate niclosamide synergism with drugs used in sporotrichosis treatment. Metabolic activity of the strains under niclosamide treatment was evaluated using the resazurin dye. Niclosamide was active against all S. brasiliensis strains (n = 17), but it was ineffective (MIC > 20 µM) for some strains (n = 4) of other pathogenic Sporothrix species. Niclosamide MIC values for Sporothrix spp. were similar for mycelial and yeast-like forms of the strains (P = 0.6604). Niclosamide was fungicidal (MFC/MIC ratio ≤ 2) for most strains studied (89%). Niclosamide activity against S. brasiliensis is independent of the fungal genotype or non-wild-type phenotypes for amphotericin B, itraconazole, or terbinafine. These antifungal drugs presented indifferent interactions with niclosamide. Niclosamide has demonstrated potential for repurposing as a treatment for sporotrichosis, particularly in S. brasiliensis cases, instigating in vivo studies to validate the in vitro findings.


Subject(s)
Anthelmintics , Antifungal Agents , Microbial Sensitivity Tests , Niclosamide , Sporothrix , Sporothrix/drug effects , Sporothrix/genetics , Sporothrix/classification , Niclosamide/pharmacology , Antifungal Agents/pharmacology , Anthelmintics/pharmacology , Sporotrichosis/microbiology , Sporotrichosis/drug therapy , Genotype , Humans , Drug Resistance, Fungal , Drug Synergism
18.
Lancet Microbe ; 5(3): e282-e290, 2024 03.
Article in English | MEDLINE | ID: mdl-38432234

ABSTRACT

BACKGROUND: Zoonotic sporotrichosis is a neglected fungal disease, whereby outbreaks are primarily driven by Sporothrix brasiliensis and linked to cat-to-human transmission. To understand the emergence and spread of sporotrichosis in Brazil, the epicentre of the current epidemic in South America, we aimed to conduct whole-genome sequencing (WGS) to describe the genomic epidemiology. METHODS: In this genomic epidemiology study, we included Sporothrix spp isolates from sporotrichosis cases from Brazil, Colombia, and the USA. We conducted WGS using Illumina NovaSeq on isolates collected by three laboratories in Brazil from humans and cats with sporotrichosis between 2013 and 2022. All isolates that were confirmed to be Sporothrix genus by internal transcribed spacer or beta-tubulin PCR sequencing were included in this study. We downloaded eight Sporothrix genome sequences from the National Center for Biotechnology Information (six from Brazil, two from Colombia). Three Sporothrix spp genome sequences from the USA were generated by the US Centers for Disease Control and Prevention as part of this study. We did phylogenetic analyses and correlated geographical and temporal case distribution with genotypic features of Sporothrix spp isolates. FINDINGS: 72 Sporothrix spp isolates from 55 human and 17 animal sporotrichosis cases were included: 67 (93%) were from Brazil, two (3%) from Colombia, and three (4%) from the USA. Cases spanned from 1999 to 2022. Most (61 [85%]) isolates were S brasiliensis, and all were reported from Brazil. Ten (14%) were Sporothrix schenckii and were reported from Brazil, USA, and Colombia. For S schenckii isolates, two distinct clades were observed wherein isolates clustered by geography. For S brasiliensis isolates, five clades separated by more than 100 000 single-nucleotide polymorphisms were observed. Among the five S brasiliensis clades, clades A and C contained isolates from both human and cat cases, and clade A contained isolates from six different states in Brazil. Compared with S brasiliensis isolates, larger genetic diversity was observed among S schenckii isolates from animal and human cases within a clade. INTERPRETATION: Our results suggest that the ongoing epidemic driven by S brasiliensis in Brazil represents several, independent emergence events followed by animal-to-animal and animal-to human transmission within and between Brazilian states. These results describe how S brasiliensis can emerge and spread within a country. FUNDING: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brazil; the São Paulo Research Foundation; Productivity in Research fellowships by the National Council for Scientific and Technological Development, and Ministry of Science and Technology of Brazil.


Subject(s)
Sporothrix , Sporotrichosis , Animals , Humans , Sporotrichosis/epidemiology , Sporotrichosis/veterinary , Sporotrichosis/microbiology , Brazil/epidemiology , Phylogeny , Disease Outbreaks , Genomics , Sporothrix/genetics
19.
Anal Chim Acta ; 1289: 342216, 2024 Feb 08.
Article in English | MEDLINE | ID: mdl-38245200

ABSTRACT

BACKGROUND: Sporothrix globosa (S. globosa) is an agricultural activity-related but neglected pathogenic fungus responsible for sporotrichosis. Timely detection is crucial for managing and preventing its spread. However, due to the lack of efficient recognition elements for enriching S. globosa, the current approaches for detecting S. globosa are not simple and/or sensitive enough. This hinders their wider application of fast screening. RESULTS: Herein, we successfully prepared immunoglobulin Y (IgY) targeting S. globosa, and developed a rapid and accurate detection method, improving upon current limited and inadequate detection approaches. Our method combined the use of IgY and loop-mediated isothermal amplification (LAMP) to enhance detection sensitivity and specificity simultaneously. The IgY was fabricated on magnetic beads to specifically concentrate S. globosa in samples, while LAMP amplified the captured target after simple boiling DNA extraction. By using our method, as low as 4.66 × 102 Cells mL-1S. globosa was accurately detected in soil and corn straw samples. We further integrated this assay into a portable toolbox for sample-to-result detection in resource-limited areas. By using this toolbox, we have colorimetrically detected soil and corn straw samples contaminated by S. globosa, suggesting the promising on-site detection potential. SIGNIFICANCE AND NOVELTY: A new IgY recognizing S. globosa was prepared. Through the combination of IgY enrichment and LAMP amplification, the detection sensitivity and specificity were improved simultaneously. This method eliminated thermal cycling, simplified the operation, and reduced the analysis time. Compared to existing methods, our approach is more suitable for on-site detection and can significantly improve public health responses to sporotrichosis outbreaks.


Subject(s)
Immunoglobulins , Molecular Diagnostic Techniques , Sporothrix , Sporotrichosis , Humans , Sporotrichosis/diagnosis , Sporotrichosis/epidemiology , Sporotrichosis/microbiology , Nucleic Acid Amplification Techniques , Sensitivity and Specificity , Soil , Magnetic Phenomena
20.
Mycoses ; 67(1): e13656, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37743555

ABSTRACT

BACKGROUND: Systemic sporotrichosis occurs when organs, other than subcutaneous tissues and lymph nodes, are infected. Interestingly, systemic sporotrichosis in apparently immunocompetent individuals is increasing in Brazil, but data on clinical manifestations and risk factors are scarce. Most of the existing data on such condition relate to people living with HIV. We aimed to study the risk factors associated with systemic sporotrichosis among HIV-negative and HIV-positive patients. METHODS: We performed a retrospective cross-sectional study with 80 patients from Brazil, diagnosed between 2014 and 2021. The association between disease classification, clinical presentation and risk factors were analysed by logistic regression. RESULTS: Of the 80 patients, 29 (36.3%) presented with systemic sporotrichosis. All HIV-positive patients developed the systemic form, with increased mortality (p = .002). Alcohol ingestion (p = .009) and diabetes (p = .010) were associated with systemic disease. Alcohol ingestion seemed to favour pulmonary infection (p = .013) and, diabetes favoured osteoarticular (p = .009) and ocular involvement (p = .033). The occurrence of fever (p = .001) and weight loss (p = .006) were significantly associated with systemic sporotrichosis, while meningeal involvement (p = .001) increased mortality risk. Nine (11.3%) patients died from sporotrichosis. The presence of fungal structures in the mycological examination of the patients' lesions were associated with the systemic form (p = .017) and death (p = .002). CONCLUSION: Our study points to the factors that drive systemic sporotrichosis other than HIV, such as alcohol ingestion and diabetes. Considering the high number of patients presenting severe sporotrichosis, the recognising these factors may contribute to timely diagnosis and proper treatment.


Subject(s)
Diabetes Mellitus , HIV Infections , Sporothrix , Sporotrichosis , Humans , Sporotrichosis/microbiology , Brazil/epidemiology , Cross-Sectional Studies , Retrospective Studies
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