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/geneticsABSTRACT
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 SpecificityABSTRACT
A 37-year-old immunocompetent man was admitted to the emergency department due to recurrent pain and oedema of his right knee. Two months earlier, he had undergone surgery to repair his meniscus. Arthroscopic joint lavage was performed and Candida dubliniensis was recovered in culture. The authors describe the first case of septic arthritis caused by Candida dubliniensis.
ABSTRACT
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.
Subject(s)
Cat Diseases , Epidemics , Sporothrix , Sporotrichosis , Cats , Animals , Humans , Sporotrichosis/epidemiology , Sporotrichosis/veterinary , Brazil/epidemiology , Disease Outbreaks/veterinary , Cat Diseases/epidemiologyABSTRACT
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.
ABSTRACT
Opportunistic infections are serious complications in critically ill COVID-19 patients, especially co-infections with bacterial and fungal agents. Here we report a rare case of bloodstream co-infection by Trichosporon asahii, an emerging yeast, and Acinetobacterbaumannii, an opportunistic nosocomial pathogen, both multidrug resistant, in a tertiary hospital from southern Brazil. A review of the literature regarding similar cases is also included. Treatment with multiple antimicrobials failed, and the patient progressed to death four days after the diagnosis of bacteremia and fungemia.
Subject(s)
COVID-19 , Coinfection , Mycoses , Sepsis , Trichosporon , Antifungal Agents/therapeutic use , Basidiomycota , COVID-19/complications , Coinfection/diagnosis , Coinfection/drug therapy , Humans , Mycoses/diagnosis , Sepsis/microbiologyABSTRACT
The checkerboard broth microdilution assay (BMD) is the most frequently used method for the in vitro evaluation of drug combinations. However, its use to evaluate the effect of antifungal drugs on filamentous fungi is sometimes associated with endpoint-reading difficulties, and different degrees of interaction are assigned to the same drug combination. We evaluated combinations of the azoles, itraconazole, posaconazole, and voriconazole, with the echinocandins, anidulafungin, caspofungin, and micafungin, against 15 itraconazole-resistant Aspergillus fumigatus clinical strains via the checkerboard BMD and Etest assay. Readings after 24 and 48 h, considering the two reading endpoints, the minimum inhibitory concentration (MIC) and minimum effective concentration (MEC), were performed for both methods. Our results showed that the correlation coefficients between the BMD and Etest methods were quite diverse to the drug combinations tested. The highest correlation coefficients of the Etest with the BMD assays (MEC and MIC reading) were the Etest-MIC reading at 24 h and the Etest-MEC reading at 48 h. Improvements in experimental conditions may increase the correlation between the two methods and ensure that Etest assay can be safely used in the evaluation of antifungal combinations against Aspergillus species.
Subject(s)
Antifungal Agents/pharmacology , Aspergillus fumigatus/drug effects , Drug Resistance, Fungal , Itraconazole/pharmacology , Microbial Sensitivity Tests/methods , Aspergillosis/microbiology , Aspergillus fumigatus/isolation & purification , Drug Interactions , Echinocandins/pharmacology , HumansABSTRACT
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.
Subject(s)
Empyema/microbiology , Lung Transplantation/adverse effects , Nocardia Infections/diagnosis , Nocardia/genetics , Empyema/diagnosis , Female , Humans , Middle Aged , ReoperationABSTRACT
Paracoccidioidomycosis (PCM) is a systemic mycosis with a geographic distribution limited to Latin America. PCM is caused by species in the genus Paracoccidioides, which usually appear in tissues as large yeasts, 5 to 30 µm in size. The daughter cells are attached to the parent cell by a narrow neck. Sometimes smaller forms occur (1 to 4 µm). These can be confused with other fungi, such as Histoplasma capsulatum and unencapsulated Cryptococcus variants. Twelve cases of PCM were reported with small forms of Paracoccidioides. The aim of this paper is to focus on the possibility of differential diagnosis with other systemic mycoses
Subject(s)
Paracoccidioidomycosis , Paracoccidioides , HistoplasmaABSTRACT
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.
Subject(s)
Humans , Female , Lung Transplantation/adverse effects , Empyema/diagnosis , Empyema/microbiology , Nocardia/genetics , Nocardia Infections/diagnosis , Reoperation , Middle AgedABSTRACT
Sarcoidosis is a multisystem disorder that is characterized by noncaseous epithelioid cell granulomas, which may affect almost any organ. Thoracic involvement is common and accounts for most of the morbidity and mortality associated with this disease. The diagnosis is based on exhaustive exclusion of differential diagnoses, particularly granulomatous infections. We report data on eight patients with paracoccidioidomycosis mimicking sarcoidosis. Five patients presented with a chronic pulmonary type infection and three had a disseminated form after immunosuppressive treatment. The mycological diagnosis in noncaseating granulomas is emphasized and reviewed.
Subject(s)
Paracoccidioidomycosis/diagnosis , Paracoccidioidomycosis/pathology , Sarcoidosis/diagnosis , Sarcoidosis/pathology , Diagnosis, Differential , Humans , Immunocompromised Host , Immunosuppressive Agents/administration & dosage , Immunosuppressive Agents/adverse effects , Paracoccidioidomycosis/microbiology , Paracoccidioidomycosis/mortality , Sarcoidosis/microbiologyABSTRACT
We describe 27 cases of fungal rhinosinusitis, which were caused by agents other than Aspergillus, diagnosed at our institution during a 24-year period. Particular focus was on defining the causal fungi and the predisposing factors. Fungal cultures were obtained from 20 cases and there was no growth in seven cases. Classification of mycotic disease of the nose and paranasal sinuses as invasive and noninvasive is based on clinical, radiological, and histopathological factors. The most common pathogens were Histoplasma capsulatum (n=4), Scedosporium apiospermum (n=2), Alternaria alternata (n=2), Schizophyllum commune (n=2), Pseudallescheria boydii (n=1), Penicillium sp. (n=1), Lichtheimia (Absidia) corymbifera (n=1), Xylaria enteroleuca (n=1), Trichoderma asperellum (n=1), T. harzianum (n=1), T. viride (n=1), Fusarium solani (n=1), Cladosporium sp. (n=1), and Cryptococcus neoformans (n=1). From the ones that revealed no growth, four were classified as hyalohyphomycosis and three were mucormycosis by the histopathological findings. In addition, we describe the first welldocumented case of rhinosinusitis and human infection by T. asperellum.
Subject(s)
Sinusitis , Aspergillus , TrichodermaABSTRACT
We describe a case of cryptococcal fungemia in a 62-year-old male renal transplant patient. The diagnosis was established by isolation of Cryptococcus neoformans using the Isolator(®) blood culture lysis-centrifugation system. Testing for cryptococcal antigens was negative in the serum and cerebrospinal fluid. Transbronchial lung biopsies and bronchoalveolar lavage were negative. Antifungal therapy with fluconazole was started, resulting in fever remission, and a sustained clinical response was achieved. The literature on miliary pulmonary cryptococcosis is reviewed, and three similar cases were previously reported with disseminated cryptococcosis that resembled miliary tuberculosis on imaging. These emphasize the importance of eliminating causes other than tuberculosis in patients presenting with miliary pulmonary disease, even in countries with a high prevalence of tuberculosis.
Subject(s)
Cryptococcosis/microbiology , Lung Diseases, Fungal/microbiology , Antifungal Agents/administration & dosage , Cryptococcosis/diagnosis , Cryptococcosis/drug therapy , Cryptococcosis/etiology , Humans , Kidney Transplantation/adverse effects , Lung Diseases, Fungal/diagnosis , Lung Diseases, Fungal/drug therapy , Lung Diseases, Fungal/etiology , Male , Middle AgedABSTRACT
Cryptococcosis, a systemic fungal infection, has become a significant, global public health problem. Patients with liver disease have an increased predisposition to infections, such as Cryptococcosis. To report the underlying disease, the variety of etiologic agents involved and the outcomes of the Cryptococcosis in patients living with HBV and/or HCV, we reviewed 34 medical records of patients who were diagnosed with Cryptococcosis by the Mycology Laboratory of Santa Casa Hospital, Porto Alegre, Brazil. Males corresponded to 79% of the patients, and the average patient age was 46.9 years. The cultures of 26/34 patients were positive: 25 patients were infected with Cryptococcus neoformans and one with C. gattii. A total of 14 deaths (41%) occurred. As a criterion of our study, all patients had viral hepatitis infection: 27 (80%) were infected with HCV, five (15%) were infected with HBV, and two patients were infected with both viruses. Because HBV and/or HCV are transmitted among drug users through infected blood, and the end-stage cirrhotic liver must be transplanted, these two population types were well represented in this study and were analyzed in detail. Cryptococcosis patients living with HCV and/or HBV appear to have the same symptoms, mean age and gender distribution as the general Cryptococcosis population. Once Cryptococcosis affects the brain, a high mortality rate ensues; therefore, physicians must be aware of the possible occurrence of this disease in patients living with HCV and HBV.
Subject(s)
Cryptococcosis/microbiology , Hepatitis B/complications , Hepatitis C/complications , Adult , Aged , Brazil/epidemiology , Cryptococcosis/epidemiology , Cryptococcosis/etiology , Cryptococcus/genetics , Cryptococcus/isolation & purification , Cryptococcus/physiology , Female , Hepatitis B/epidemiology , Hepatitis C/epidemiology , Humans , Male , Middle AgedABSTRACT
Aspergillus species are considered opportunistic fungi of increasing clinical importance. Informationregarding extrapulmonary involvement is scarce. The aim of this study was to isolate the differentspecies of Aspergillus from patients with rhinosinusitis. A retrospective study was conducted ina university hospital in Porto Alegre, Brazil (19862014). For mycological diagnoses, paranasaltissue obtained at surgery was subjected to histopathology examination and sent for fungal cultures.Of the 54 samples analyzed, 32 were diagnosed positive by culture. The underlying causes ofimmunodeficiency were: six with transplantation (three bone marrow,two lung, one kidney) andtwo with hematological disease (one bone marrow neoplasia and two leukemia). In the presentstudy, the clinical manifestations of rhinosinusitis aspergillosis were: 20 allergic reactions, 20fungus balls, and 14 acute invasive cases. The species isolated from the 54 samples were: Aspergillusfumigatus (n=14); A. flavus (n=6); A. niger (n=2); A. terreus (n=1); A. fischeri (n=1); and Aspergillussp., (n=3). Two concomitant species of Aspergillus were observed in two patients: A. fumigatus andA. flavus; and A. fumigatus and A. niger. In four patients, Aspergillus was associated with other fungi. These were: A. flavus and Fusarium, A. fumigatus and Rhyzopus, A. flavus and Mucorales, and Aspergillus sp. and Mucorales. The most common species of Aspergillus that were responsiblefor paranasal sinus infections were A. fumigatus, A. flavus, and A. niger...
Espécies de Aspergillus são considerados fungos oportunistas de crescente importância clínica.Informações sobre o envolvimento extrapulmonar é escassa. O objetivo deste estudo foi isolaras diferentes espécies de Aspergillus em pacientes com rinossinusite. Um estudo retrospectivofoi realizado em um hospital universitário em Porto Alegre, Brasil (1986-2014). Para diagnósticomicológico, tecido paranasais obtido no momento da cirurgia foi submetido a exame histopatológicoe encaminhados para cultivos de fungos. Das 54 amostras analisadas, 32 foram diagnosticados pelocultivo positivo. As causas subjacentes da imunodeficiência foram: seis com transplante (medulaóssea, três, pulmão, dois; rim, um) e dois com doenças hematológicas (neoplasia osso estreito,um; leucemia, duas). No presente estudo, as manifestações clínicas de rinossinusite aspergilarforam: alérgica, 20; bolas fúngica, 20; e aguda invasiva, 14. As espécies fúngicas isoladas foram:Aspergillus fumigatus, 14; A. flavus, seis; A. niger, dois; A. terreus, um; A. fischeri, um; e Aspergillussp., três. Duas espécies de Aspergillus concomitantes foram observadas em dois pacientes: A.fumigatus e A. flavus; e A. fumigatus e A. niger. Em quatro pacientes, Aspergillus foi associado comoutros fungos: A. flavus e Fusarium, um; A. fumigatus e Rhyzopus, um; A. flavus e Mucorales, um; eAspergillus sp. e Mucorales, um. Os isolados mais comuns de Aspergillus que são responsáveis porinfecções dos seios paranasais são A. fumigatus, A. flavus e A. niger...
Subject(s)
Humans , Aspergillus flavus/isolation & purification , Aspergillus fumigatus/isolation & purification , Aspergillus niger/isolation & purification , Aspergillosis , Aspergillosis, Allergic BronchopulmonaryABSTRACT
Candida albicans is often isolated from clinical samples, thus its presumptive differentiation from other species of the same genus can be based on its ability to form the germ tube in human serum. Nevertheless, there are two other species that share this characteristic: C. dubliniensis and C. africana. The aim of this study was to compare four different substrates to perform the germ tube (GT) test. The Candida spp. isolates were identified using a manual system (135 C. albicans, 24 C. tropicalis and one C. dubliniensis). The germ tube test was performed with fresh, previously frozen serum and Mueller-Hinton (MH) broth and agar. GT was observed in 96% (130/136) of the isolates through the fresh serum technique, 94% (128/136) through previously frozen serum, 92% (125/136) in MH agar, and 90% (122/136) in MH broth. The sensitivity of each test was higher than 90%, with 100% specificity. Both the MH agar and broth were able to identify the true positives, and false positives were not found. However, some C. albicans isolates were not identified. MH agar and broth may be used in laboratory for the rapid presumptive identification of C. albicans, as an alternative method for germ tube test.
Candida albicans é frequentemente isolada em amostras clínicas, assim a sua diferenciação presuntiva de outras espécies do gênero pode ser baseada na habilidade em formar o tubo germinativo em soro humano. Entretanto, existem outras duas espécies que também possuem essa característica, C. dubliniensis e C. africana. O objetivo foi comparar quatro diferentes substratos para a realização da prova do tubo germinativo (TG). Utilizou-se isolados de Candida spp. identificados através de meio manual (135 C. albicans, 24 C. tropicalis e um C. dubliniensis). A prova do tubo germinativo foi realizada utilizando soro previamente congelado e fresco, caldo e ágar Mueller-Hinton (MH). O TG através da técnica do soro a fresco foi observado em 96% (130/136), 94% (128/136) através do soro previamente congelado, 92% (125/136) no ágar e 90% (122/136) no caldo MH. A sensibilidade de cada teste foi maior que 90% e especificidade de 100%. Tanto o caldo quanto o ágar MH foram capazes de identificar apenas os verdadeiros positivos e não ocorrendo falsos positivos, porém deixaram de identificar alguns isolados de C. albicans. O ágar e o caldo MH podem ser utilizados na rápida e presuntiva identificação laboratorial de C. albicans, como uma alternativa para o teste do tubo germinativo.
Subject(s)
Humans , Agar/pharmacology , Candida/classification , Culture Media/chemistry , Mycological Typing Techniques/methods , Candida/growth & development , Sensitivity and Specificity , Species SpecificityABSTRACT
Candida albicans is often isolated from clinical samples, thus its presumptive differentiation from other species of the same genus can be based on its ability to form the germ tube in human serum. Nevertheless, there are two other species that share this characteristic: C. dubliniensis and C. africana. The aim of this study was to compare four different substrates to perform the germ tube (GT) test. The Candida spp. isolates were identified using a manual system (135 C. albicans, 24 C. tropicalis and one C. dubliniensis). The germ tube test was performed with fresh, previously frozen serum and Mueller-Hinton (MH) broth and agar. GT was observed in 96% (130/136) of the isolates through the fresh serum technique, 94% (128/136) through previously frozen serum, 92% (125/136) in MH agar, and 90% (122/136) in MH broth. The sensitivity of each test was higher than 90%, with 100% specificity. Both the MH agar and broth were able to identify the true positives, and false positives were not found. However, some C. albicans isolates were not identified. MH agar and broth may be used in laboratory for the rapid presumptive identification of C. albicans, as an alternative method for germ tube test.
Subject(s)
Agar/pharmacology , Candida/classification , Culture Media/chemistry , Mycological Typing Techniques/methods , Candida/growth & development , Humans , Sensitivity and Specificity , Species SpecificityABSTRACT
Nondiphtherial corynebacteria are ubiquitous in nature and commonly colonize the skin and mucous membranes of humans, however they rarely account for clinical infection. We present the first reported case of multiple pulmonary nodules caused by Corynebacterium striatum. The infection occurred in a 72-year-old immunocompetent female, and the diagnosis was obtained by Gram's stain and culture of lung biopsy. C. striatum should be recognized as a potential pathogen in both immunocompromised and normal hosts in the appropriate circumstances.
Subject(s)
Corynebacterium Infections/diagnosis , Multiple Pulmonary Nodules/microbiology , Aged , Corynebacterium Infections/microbiology , Female , Humans , Immunocompetence , Multiple Pulmonary Nodules/diagnosisABSTRACT
Nondiphtherial corynebacteria are ubiquitous in nature and commonly colonize the skin and mucous membranes of humans, however they rarely account for clinical infection. We present the first reported case of multiple pulmonary nodules caused by Corynebacterium striatum. The infection occurred in a 72-year-old immunocompetent female, and the diagnosis was obtained by Gram's stain and culture of lung biopsy. C. striatum should be recognized as a potential pathogen in both immunocompromised and normal hosts in the appropriate circumstances.
Bacilos não diftéricos são ubiquitários na natureza e comumente colonizam a pele e as membranas mucosas humanas, contudo eles raramente acarretam doença clínica. Apresentamos o primeiro relato de múltiplos nódulos causados por Corynebacterium striatum. A infecção ocorreu numa mulher imunocompetente de 72 anos de idade e o diagnóstico foi obtido pela coloração de Gram e cultivo de biópsia pulmonar. C. striatum deve ser reconhecido como potencial patógeno tanto em pacientes imunodeprimidos como em hospedeiros normais, em circunstâncias apropriadas.
Subject(s)
Aged , Female , Humans , Corynebacterium Infections/diagnosis , Multiple Pulmonary Nodules/microbiology , Corynebacterium Infections/microbiology , Immunocompetence , Multiple Pulmonary Nodules/diagnosisABSTRACT
Cryptococcus gattii are closely related species of encapsulated yeast-like fungi involved in the etiology of cryptococcosis, especially in immunocompetent individuals. Dissemination with involvement of many organ systems is common. On the other hand, cellulitis in an immunossupressed patient caused by C. gattii is rare. We present a case of disseminated disease caused by Cryptococcus gattii in a lung transplant recipient who manifested cellulitis. The disease was also complicated by a lung carcinoma. We emphasize that cryptococcal cellulitis related to C. gattii in solid organ transplant (SOT) recipients should be considered in the differential diagnosis of acute bacterial skin infections.