Riesgo patogénico por Aspergillus fumigatus en la producción de compost de Musa paradisiaca / Pathogenic risk by Aspergillus fumigatus in the compost production of Musa paradisiaca

Aspergillus fumigatus es un hongo con características de saprofito que crece en las partes muertas y en descomposición de la materia orgánica. La presencia de terapias inmunosupresoras o de antifúngicos pueden convertirlo en un patógeno fúngico oportunista causante de enfermedades como aspergilosis pulmonar crónica (CPA), aspergilosis invasiva aguda y síndrome de alergias. Pacientes que sufren de enfermedad pulmonar severa como la pulmonar obstructiva crónica (EPOC) tienen un mayor riesgo de desarrollar aspergilosis pulmonar crónica y aspergilosis pulmonar necrosante crónica (CNPA) que pueden causar crecimiento de hongos en los tejidos dañados. Por otro lado, una de las fuentes más comunes de estos hongos son la generación de compost. Estos fertilizantes orgánicos no dañan al medio ambiente y podrían resultar una estrategia eficaz para salvaguardar el medio ambiente. El compost orgánico promueve el suelo con mejor capacidad de retención de agua y resistencia a la sequía. Entre los residuos agrícolas postcosecha más comunes utilizados en la formación de compost, se pueden mencionar a las cáscaras de yuca, de maní, de frutas y verduras, desechos de cervecería, cascarilla de cacao, cascarilla de cola, maíz, conchas de plátano (Musa paradisiaca). Esta investigación tuvo por objetivo determinar la calidad del compost producido a partir de la cáscara de plátano con el fin de aprovechar hasta un 70% de su biomasa en la Municipalidad Provincial de Leoncio Prado, Perú. Los resultados mostraron que la la degradación aerobia de la cáscara de M. paradisiaca generó emisiones de CO2 (88,61%), así como compost (6,57 %) y lixiviados (4,82 %). Sin embargo, los efectos fitotóxicos usando el índice de germinación mostraron una inhibición completa de las semillas probadas posiblemente debido al gran contenido de minerales (alto contenido de salinidad del compost y lixiviado), presencia de metales pesados, compuestos orgánicos presentes, o la presencia de hongos tales como el A. fumigatus(AU)

Aspergillus fumigatus is a saprophytic fungus that grows on dead and decomposing parts of organic matter. The presence of immunosuppressive or antifungal therapies can make it an opportunistic fungal pathogen causing diseases such as chronic pulmonary aspergillosis (CPA), acute invasive aspergillosis, and allergy syndrome. Patients suffering from severe lung disease such as chronic obstructive pulmonary disease (COPD) are at increased risk of developing chronic pulmonary aspergillosis and chronic necrotizing pulmonary aspergillosis (CNPA) which can cause fungal growth in damaged tissues. On the other hand, one of the most common sources of these fungi is the generation of compost. These organic fertilizers do not harm the environment and could be an effective strategy to safeguard the environment. Organic compost promotes soil with better water holding capacity and drought resistance. Among the most common postharvest agricultural residues used in the formation of compost, we can mention cassava shells, peanut shells, fruit and vegetable shells, brewery waste, cocoa shells, tail shells, corn, banana shells (Musa paradisiaca). The objective of this research was to determine the quality of the compost produced from the banana peel in order to take advantage of up to 70% of its biomass in the Provincial Municipality of Leoncio Prado, Peru. The results showed that the aerobic degradation of the shell of M. paradisiaca generated CO2 emissions (88.61%), as well as compost (6.57%) and leachates (4.82%). However, the phytotoxic effects using the germination index showed a complete inhibition of the tested seeds, possibly due to the high mineral content (high salinity content of the compost and leachate), the presence of heavy metals, organic compounds present, or the presence of fungi such as A. fumigatus(AU)

Conocimientos y tendencias actuales sobre la aspergilosis broncopulmonar alérgica / Current knowledge and trends on allergic bronchopulmonary aspergillosis

Introducción: La aspergilosis broncopulmonar alérgica es una enfermedad pulmonar inflamatoria, caracterizada por una reacción de hipersensibilidad a la colonización de la vía aérea por Aspergillus fumigatus en individuos susceptibles, generalmente pacientes con asma y fibrosis quística. Objetivo: Actualizar los conocimientos en el diagnóstico y manejo de la aspergilosis broncopulmonar alérgica. Desarrollo: Se realizó una revisión bibliográfica temática, observacional y retrospectiva, de diciembre de 2019 a julio de 2020. Se accedió a bases de datos como PubMed/MEDLINE, SciElo, ReserchGate, Scopus, ScienceDirect, Redalyc, Latindex, Wed of Science y LILACS, usando los descriptores en Ciencias de la Salud (Aspergillus fumigatus, aspergilosis broncopulmonar alérgica, hipersensibilidad, allergic bronchopulmonary aspergillosis, aspergilose broncopulmonar alérgica, pulmonary aspergillosis, aspergilose pulmonar, hypersensitivity, hipersensibilidade). Se incluyeron artículos escritos en español, inglés o portugués, que informaron sobre la temática relacionada con aspergilosis broncopulmonar alérgica; seleccionados por la lectura del título, el resumen, el texto completo y por la evaluación de las variables (tipos de diseño metodológico, aspectos metodológicos generales propios de cada uno de los diseños encontrados y calidad de la investigación). Con estos criterios, fueron seleccionados 36 artículos nacionales e internacionales que incluyeron artículos originales, revisiones bibliográficas, estudios de intervención, revistas y anuarios estadísticos. Conclusiones: La aspergilosis broncopulmonar alérgica es mucho más frecuente de lo que se suponía, pero con asiduidad se subdiagnostica y el diagnóstico se hace tardíamente, lo que empeora el pronóstico de estos pacientes. Establecer el diagnóstico y tratamiento oportuno ayudará prevenir el desarrollo de serias complicaciones(AU)

Introduction: Allergic bronchopulmonary aspergillosis is an inflammatory lung disease characterized by a hypersensitivity reaction to colonization of the airway by Aspergillus fumigatus in susceptible individuals, generally patients with asthma and cystic fibrosis. Objective: To update knowledge in the diagnosis and management of allergic bronchopulmonary aspergillosis. Development: A thematic, observational and retrospective bibliographic review was carried out, from December / 2019 to July / 2020. Databases such as PubMed / MEDLINE, SciElo, ReserchGate, Scopus, ScienceDirect, Redalyc, Latindex, Wed of Science and LILACS were accessed, using descriptors in Health Sciences (Aspergillus fumigatus, allergic bronchopulmonary aspergillosis, hypersensitivity, allergic bronchopulmonary aspergillosis, allergic bronchopulmonary aspergillose, pulmonary aspergillosis, pulmonary aspergillus, hypersensitivity, hypersensitivity). Articles written in Spanish, English or Portuguese were included, which reported on the subject related to allergic bronchopulmonary aspergillosis, selected by reading the title, the abstract, the full text and by evaluating the variables (types of methodological design, aspects general methodological characteristics of each of the designs found and the quality of the research). With these criteria, 36 national and international articles were selected that included original articles, bibliographic reviews, intervention studies, journals and statistical yearbooks. Conclusions: Allergic bronchopulmonary aspergillosis is much more frequent than previously assumed, but it is frequently underdiagnosed and the diagnosis is made late, which worsens the prognosis of these patients. Establishing timely diagnosis and treatment will help prevent the development of serious complications(AU)

Prevalencia de aspergilosis broncopulmonar alérgica en niños con fibrosis quística / Prevalence of bronchopulmonary aspergillosis in children with cystic fibrosis

La aspergilosis broncopulmonar alérgica (ABPA) es una reacción de hipersensibilidad secundaria al Aspergillus fumigatus (Af) que complica la evolución en fibrosis quística (FQ). Existen pocos estudios pediátricos de su prevalencia publicados en el mundo y en Chile se desconoce. El objetivo de este trabajo fue estimar la prevalencia de ABPA en niños con FQ en un hospital de referencia, explorar factores de riesgo y describir los criterios diagnósticos, tratamiento y evolución. Se incluyeron retrospectivamente los niños con FQ atendidos en un hospital terciario en Santiago de Chile (Hospital Roberto del Río) entre los años 2011 a 2019, se identificaron aquellos con diagnóstico de ABPA. Se registraron criterios diagnósticos según la Cystic Fibrosis Foundation, presencia de factores de riesgo, tratamientos recibidos y efectos adversos. De 65 pacientes con FQ atendidos en este período, la prevalencia de ABPA fue del 12%. El promedio de edad al diagnóstico fue ± 11 años (5-17 años), predominando la edad adolescente y el género masculino. El 50% cumplieron con los criterios clásicos, el 87,5% usaron antibióticos y el 62,5% corticoides inhalados. La respuesta favorable al tratamiento inicial con corticoides y antifúngico vía oral fue 62,5%, con una exacerbación al momento del estudio. El 25% se comportaron como refractario y el 12,5% respondieron a tratamiento con pulsos de metilprednisolona. El 37,5% presentaron eventos adversos relacionados a corticoides. La prevalencia de ABPA observada es comparable a las series publicadas. Se necesitan trabajos prospectivos para conocer la prevalencia nacional y su tendencia a lo largo de los años, identificando factores de riesgo.

Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity response to Aspergillus fumigatus (Af) and worsens outcome in children with cystic fibrosis (CF). Its prevalence varies in the literature, but we do not know it in Chile. The aim of the study was to know the prevalence of ABPA in children with CF and to describe risk factors, diagnostic criteria, treatment and outcome. We included all patients with CF seen in a tertiary hospital in Santiago, Chile (Hospital Roberto del Río), between 2011 and 2019; ABPA cases (CF Foundation diagnostic criteria) were identified for the estimation of the prevalence. Risk factors, diagnostic criteria and treatment were recorded, as proposed by the Cystic Fibrosis Foundation. A total of 65 patients with CF were identified in the study period, with a prevalence of 12% (8 cases). Mean age at diagnosis ± 11 years (5-17), more frequent in adolescence and male. CF Foundation criteria diagnostic were identified in 50% of cases, with high frequency of antibiotic use (87,5%) and inhaled steroids (62,5%). Positive oral steroids and antifungal treatment response was 62,5%. Refractary response was 25% and 12,5% needed intravenous metilprednisolone pulses. A 37,5% of cases presented adverse effects to steroids. Prevalence of ABPA is comparable to literature. A prospective study is needed to identified national prevalence and trends, identifying risks factors.

Outbreak of avian aspergillosis in colonial-bred chicks (Isa Brown) in southern Rio Grande do Sul - case report / [Surto de aspergilose aviária em pintainhas (Isa Brown) de criação colonial na região sul do Rio Grande do Sul - relato de caso]

In this study we describe the epidemiology, clinical signs, and pathology of an outbreak of avian aspergillosis in alternative breeding in the southern region of Rio Grande do Sul, Brazil. Between the fifth and tenth day of life, 360 chicks from a flock of 4000 developed unspecific clinical signs and died. The birds were housed in a reused aviary litter, without previous treatment. In 11 six-day-old female ISA Brown chicks (Gallus gallus domesticus), necropsy revealed firm, yellowish-white, multinodular lesions extending from the pleura to the lung parenchyma. Histologically, a granulomatous, multifocal to coalescent pneumonia was observed. Granulomas were characterized by central necrosis, with heterophil and epithelioid macrophage infiltration and presence of countless Y-shaped intralesional septate hyphae morphologically compatible with Aspergillus spp. The diagnosis through isolation confirmed Aspergillus fumigatus. We highlight the importance of aspergillosis as a primary cause of diseases in the respiratory tract of young birds in alternative breeding. Measures to prevent aspergillosis mainly regarding the reuse of aviary litter are essential in poultry husbandry to prevent economic losses, reduce environmental contamination and mitigate the potential risk to public health.(AU)

Descrevem-se os aspectos epidemiológicos e patológicos de um surto de aspergilose aviária em criação alternativa na região sul do Rio Grande do Sul, Brasil. De um lote de 4000 pintainhas, entre o quinto e o 10º dia de vida, 360 aves apresentaram sinais clínicos inespecíficos e morreram. As aves foram alojadas em cama reutilizada do aviário, sem tratamento prévio. Na necropsia de 11 pintainhas (Gallus gallus domesticus), fêmeas, seis dias de idade da linhagem Isa Brown, foram observadas no pulmão lesões multinodulares, branco-amareladas e firmes, que se estendiam da pleura ao parênquima. Histologicamente foi observada pneumonia granulomatosa, multifocal a coalescente. Os granulomas eram caracterizados por necrose central, com infiltrado inflamatório de heterófilos, macrófagos, células epitelioides com presença de inúmeras hifas septadas intralesionais, semelhantes à letra "Y", morfologicamente compatíveis com Aspergillus spp. O diagnóstico foi confirmado pelo isolamento de Aspergillus fumigatus. Alerta-se para a importância da aspergilose como causa primária de afecções no trato respiratório de aves jovens em criações alternativas. Medidas preventivas relacionadas ao manejo dessas aves são indispensáveis principalmente quanto à reutilização da cama dos aviários, a fim de evitar perdas econômicas, reduzir a contaminação ambiental e o potencial risco à saúde pública.(AU)

Antifungal susceptibility profile of Aspergillus fumigatus isolates from avian lungs / Perfil de suscetibilidade antifúngica de isolados de Aspergillus fumigatus provenientes de pulmões de aves

Susceptibility testing is essential to inform the correct management of Aspergillus infections. In this study we present antifungal susceptibility profile of A. fumigatus isolates recovered from lungs of birds with and without aspergillosis. Fifty three isolates were tested for their antifungal susceptibility to voriconazole (VRC), itraconazole (ITZ), amphotericin (AMB) and caspofungin (CSP) using the M38-A2 broth microdilution reference method. Five isolates were resistant to more than one antifungal drug (CSP + AMB, VRC + ITZ and AMB + ITZ). Fifteen (28%) isolates with susceptible increased exposure (I) to ITZ were sensible to VRC. Resistance to AMB (>2µg/mL) was observed in only four isolates. Eleven (21%) A. fumigatus present resistance to ITZ (13%) and VRC (8%). Fungal isolation from respiratory samples has been regarded as being of limited usefulness in the ante mortem diagnosis of aspergillosis in birds. However, the results suggest that the detection and antifungal susceptibility profile may be helpful for monitoring of therapy for avian species and where antifungal resistance might be emerging and what conditions are associated to the event.(AU)

Os testes de suscetibilidade são essenciais para informar o correto manejo das infecções por Aspergillus. Neste estudo apresentamos o perfil antifúngico de isolados de A. fumigatus provenientes de pulmões de aves com e sem aspergilose. Cinqüenta e três isolados foram testados quanto à susceptibilidade antifúngica ao voriconazol (VRC), itraconazol (ITZ), anfotericina B (AMB) e caspofungina (CSP) pelo método de referência de microdiluição do caldo M38-A2. Cinco isolados foram resistentes a mais de um antifúngico (CSP + AMB, VRC + ITZ e AMB + ITZ). Quinze (28%) isolados suscetíveis - com exposição aumentada (I) ao ITZ foram sensíveis ao VRC. A resistência ao AMB (>2µg/mL) foi observada em apenas quatro isolados. Onze (21%) A. fumigatus apresentaram resistência a ITZ (13%) e VRC (8%). O isolamento de fungos de amostras respiratórias tem sido considerado de utilidade limitada no diagnóstico ante mortem de aspergilose em aves. No entanto, os resultados sugerem que a detecção e o perfil de suscetibilidade a antifúngicos podem ser úteis para o monitoramento da terapia de espécies aviárias, assim como a emergência da resistência antifúngica e quais condições podem estar associadas ao evento.(AU)

Ellagic acid production using polyphenols from orange peel waste by submerged fermentation

Background: Biotechnological processes are part of modern industry as well as stricter environmental requirements. The need to reduce production costs and pollution demands for alternatives that involve the integral use of agro-industrial waste to produce bioactive compounds. The citrus industry generates large amounts of wastes due to the destruction of the fruits by microorganisms and insects together with the large amounts of orange waste generated during the production of juice and for sale fresh. The aim of this study was used orange wastes rich in polyphenolic compounds can be used as source carbon of Aspergillus fumigatus MUM 1603 to generate high added value compounds, for example, ellagic acid and other molecules of polyphenolic origin through submerged fermentation system. Results: The orange peel waste had a high concentration of polyphenols, 28% being condensed, 27% ellagitannins, 25% flavonoids and 20% gallotannins. The major polyphenolic compounds were catechin, EA and quercetin. The conditions, using an experimental design of central compounds, that allow the production of the maximum concentration of EA (18.68 mg/g) were found to be: temperature 30°C, inoculum 2 × 107 (spores/g) and orange peel polyphenols 6.2 (g/L). Conclusion: The submerged fermentation process is an effective methodology for the biotransformation of molecules present in orange waste to obtain high value-added as ellagic acid that can be used as powerful antioxidants, antibacterial and other applications.

Actividad antimicrobiana del aceite esencial de hojas de Psidium cattleianum Afzel. ex Sabine / Antimicrobial activity of essential oil from Psidium cattleianum Afzel. ex Sabine leaves

La búsqueda de fuentes naturales para controlar los microorganismos es de interés en la producción de alimentos. Este estudio evaluó la composición química y la actividad antimicrobiana del aceite esencial de las hojas de Psidium cattleianum. El aceite esencial se extrajo por hidrodestilación y se identificó por GC-MS. La clase predominante de compuestos fueron los sesquiterpenos (47,6%) y los principales fueron trans-ß-cariofileno (14,7%), 1,8-cineol (11,7%) y É£-muuroleno (5,6%). Actividad antimicrobiana se realizó mediante la técnica de microdilución contra ocho hongos y ocho bacterias. Concentración inhibitoria mínima varió de 0,17 a 11,25 mg mL-1 para hongos y de 1,40 a 16,87 mg mL-1 para bacterias. Principales actividades fueron contra hongos Aspergillus fumigatus (ATCC 1022), Aspergillus ochraceus (ATCC 12066), Aspergillus versicolor (ATCC 11730) y Trichoderma viride (IAM 5061), y bacterias Pseudomonas aeruginosa (ATCC 27853), Bacillus cereus (clinical isolate) y Staphylococcus aureus (ATCC 6538) con potencial para prevenir enfermedades transmitidas por alimentos.

The search for natural sources to control microorganisms is of interest in food production. This study evaluated the chemical composition and antimicrobial activity of the essential oil from Psidium cattleianum leaves. The essential oil was extracted by hydrodistillation, and identified by GC-MS. The predominant class of compounds was sesquiterpenes (47.6%) and the major compounds were trans-ß-caryophyllene (14.7%), 1,8-cineole (11.7%) and É£-muurolene (5.6%). The antimicrobial activity was carried out by microdillution technique against eight fungi and eight bacteria. The minimum inhibitory concentration ranged from 0.17 to 11.25 mg mL-1 for fungi, and from 1.40 to 16.87 mg mL-1 for bacteria. The highest activities were against fungi Aspergillus fumigatus (ATCC 1022), Aspergillus ochraceus (ATCC 12066), Aspergillus versicolor (ATCC 11730), and Trichoderma viride (IAM 5061), and bacteria Pseudomonas aeruginosa (ATCC 27853), Bacillus cereus (clinical isolate), and Staphylococcus aureus (ATCC 6538) with potential to prevent foodborne diseases.

Wide diversity of fungal species found in wellwater for human consumption: an analytical cross-sectional study

ABSTRACT BACKGROUND: Fungi are ubiquitous in the environment. They are able to grow in water and many of them may be opportunistic pathogens. OBJECTIVE: The aims were to identify fungi in registered wells (RWs) and nonregistered wells (NRWs) that tap into groundwater; and to correlate the results from physicochemical assays on this water (free residual chlorine and pH) with the presence of fungi. DATA AND SETTING: Analytical cross-sectional quantitative study on groundwater wells in São José do Rio Preto, São Paulo, Brazil. METHODS: 52 samples of 500 ml of water were collected from RWs and 107 from NRWs. These were sent to a microbiology laboratory to identify any fungi that were present. In addition, free residual chlorine and pH were measured immediately after sample collection. Several statistical analysis tests were used. RESULTS: Fungal contamination was present in 78.8% of the samples from RWs and 81.3% from NRWs. Filamentous fungi were more prevalent than yeast in both types of wells. There was no significant difference in presence of fungi according to whether chloride and pH were within recommended levels in RWs; or according to whether pH was within recommended levels in NRWs. Furthermore, there was no statistical difference in the levels of fungal contamination between RWs and NRWs. CONCLUSION: Both RWs and NRWs are potential reservoirs for many types of fungi. Many of these may become opportunistic pathogens if they infect immunosuppressed individuals. Furthermore, this study confirms that fungi are able to grow even when chlorine and pH parameters are within the standards recommended.

Identificación molecular de Aspergillus fumigatus aislados de pacientes con aspergilosis invasiva / Molecular identification of Aspergillus fumigatus isolated from patients with invasive aspergillosis

RESUMEN El objetivo del estudio fue identificar molecularmente cepas de aspergillus aislados de pacientes con aspergilosis invasiva (AI), que fueron tipificadas primariamente como Aspergillus fumigatus sensu lato por métodos fenotípicos convencionales. Se trabajó con 20 cepas de la micoteca de la sección de micología del Instituto de Medicina Tropical "Daniel A. Carrión". Para obtener el ADN fúngico se emplearon las técnicas de choque térmico, tratamiento enzimático y columnas de silica-gel; y se almacenó a -20 0C para conservarlo. En el procedimiento de la reacción en cadena de la polimerasa en tiempo real (qPCR) se incluyeron primers marcados con fluorocromo, los cuales amplificaron las secuencias específicas de A. fumigatus. La fluorescencia se midió con el termociclador al final de la fase de hibridación de cada ciclo. Se identificó molecularmente que sólo el 50% de las cepas estudiadas pertenecen a la especie Aspergillus fumigatus sensu stricto.

ABSTRACT The objective of the study was to identify molecularly-isolated strains of Aspergillus from patients with invasive aspergillosis (IA); these strains were primarily typed as Aspergillus fumigatus sensu lato by conventional phenotypic methods. We worked with 20 strains from the mycology section of the Institute of Tropical Medicine "Daniel A. Carrión." To obtain the fungal DNA, thermal shock, enzymatic treatment, and silica gel column techniques were used; and it was stored at -20°C to preserve it. The real-time polymerase chain reaction (qPCR) procedure included fluorochrome-labeled primers, which amplified the specific sequences of A. fumigatus. Fluorescence was measured with the thermocycler at the end of the hybridization phase of each cycle. It was molecularly-identified that only 50% of the strains studied belong to the species Aspergillus fumigatus sensu stricto.

Drug-induced toxic epidermal necrolysis with secondary aspergillus fumigatus infection: a case report / 北京大学学报(医学版)

Among the various drug induced dermatological entities toxic epidermalnecrolysis (TEN) and Stevens-Johnson syndrome (SJS) occupy a primary place in terms of mortality. Toxic epidermal necrolysis also known as Lyell's syndrome was first described by Lyell in 1956. Drugs are by far the most common cause of toxic epidermal necrolysis, in which large sheets of skin are lost from the body surface making redundant the barrier function of the skin, with its resultant complications. Drug-induced toxic epidermal necrolysis are severe adverse cutaneous drug reactions to various precipitating agents that predominantly involve the skin and mucous membranes. Toxic epidermal necrolysis is rare but considered medical emergencies as they are potentially fatal. Drugs are the most common cause accounting for about 65%-80% of the cases. The most common offending agents are sulfonamides, NSAIDs, butazones and hydrantoins. An immune mechanism is implicated in the pathogenesis, but its nature is still unclear. There is a prodormal phase in which there is burning sensation all over the skin and conjunctivae, along with skin tenderness, fever, malaise and arthralgias. Early sites of cutaneous involvement are the presternal region of the trunk and the face, but also the palms and soles, rapidly spread to their maximum extent, the oral mucosa and conjunctiva being affected. Initial lesions are macular, followed by desquamateion, or may be from atypical targets with purpuriccenters that coalesce, from bullae, then slough. The earlier a causative agent is withdrawn the better is the prognosis. Several treatment modalities given in addition to supportive care are reported in the literature, such as systemicsteroids, high-dose intravenous immunoglobulins, ciclosporin, TNF antagonists. Recovery is slow over a period of 14-28 days and relapses are frequent. Mortality is 25%-50% and half the deaths occur due to secondary infection. Here we report a 50-year-old female of drug-induced toxic epidermal necrolysis. She was admitted to the dermatology ward with extensive peeling of skin over the trunk and limbs. She had taken alamotrigine for epilepsy. A week after taking the tablets, the patient developed a severe burning sensation all over the body and followed by a polymorphic erythematous dermatitis and widespread peeling of skin. We treated this patient with high dose corticosteroids, high-dose intravenous immunoglobulins and etanercept, but eventually she died of secondary aspergillus fumigatus infection.

Development of Aspergillus fumigatus-induced chronic atopic dermatitis mouse model

PURPOSE: Atopic dermatitis (AD) is the most common chronic and relapsing inflammatory skin disease with skin barrier defects and altered immune responses. Chronic inflammation leads to irreversible fibrosis in the skin and there is no treatment to completely abolish the inflammation and fibrosis. To prevent or treat the chronic process of AD, it is necessary to develop a murine model of AD that reflects the chronic process to identify the mechanism. The aims of this study were to develop a chronic AD model with a crude extract Aspergillus fumigatus (Af) antigen. METHODS: We applied Af extract (40 µg) epicutaneously to the dorsal skin of BALB/c mice for 5 consecutive days per week during a period of 5 weeks for a chronic AD model, and 5 consecutive days repeatedly with 2 weeks interval for an acute AD model. RESULTS: The clinical score and transepidermal water loss were more increased in the chronic AD model than in the acute AD model. Histologic findings showed that more increased epidermal thickness, neutrophil infiltration and hyperkeratosis in the chronic model than in the acute model. Skin fibrosis was more prominent in the chronic model than in the acute model. The mRNA expression levels of transforming growth factor (TGF)-β, thymic stromal lymphopoietin, and interleukin-33 were increased in the skin of the chronic model compared to the acute model. The levels of total IgE, Af-specific IgE, IgG1, and IgG2a were significantly increased in the chronic model compared to controls. CONCLUSION: The Af-induced chronic AD model showed prominent fibrosis and increased TGF-β expression in the skin, which suggests that these models may be useful in the research for the mechanism of the chronic process in AD.

Fungicidas agrícolas azólicos e pressão seletiva de fenótipos resistentes em Aspergillus fumigatus / Agricultural azole fungicides and selective pressure of resistant phenotypes in Aspergillus fumigatus

A utilização de fungicidas azólicos pode estar ligada a resistência aos fármacos utilizados no tratamento da aspergilose invasiva. Em cepas de Aspergillus fumigatus, foram descritas mutações no gene cyp51A resultando em menor afinidade azol-alvo e resistência antifúngica. Mecanismos de resistência aos triazóis já foram descritos em isolados ambientais de áreas agrícolas e em pacientes sob tratamento. A similaridade dos compostos azólicos, clínicos e agrícolas, e o mesmo mecanismo de ação na célula fúngica podem contribuir para a seleção de cepas com resistência cruzada no hospedeiro e no meio ambiente. O gene cyp51A codifica a enzima-chave na produção de ergosterol da membrana celular-14-demetilase, alvo de todos os compostos azólicos. Mutações no gene cyp51A resultam na produção de enzima com alterações em sua conformação tridimensional levando à menor afinidade azol-alvo e resistência antifúngica. A indução de tais mutações foi atribuída à exposição extensiva das cepas ambientais a agrotóxicos fungicidas triazólicos. Estudos em diversos países sugerem que esses produtos exercem pressão seletiva em fenótipos resistentes de A. fumigatus O objetivo deste estudo foi avaliar a hipótese da origem ambiental de mecanismos de resistência a triazólicos, em 2 isolados clínicos e 2 ambientais de A. fumigatus, modulada pela exposição a um fungicida agrícola. Modelo de horta experimental, parte orgânica e parte exposta a agrotóxico triazólico, foi construída e inoculada com os 4 isolados, todos selvagens não mutantes de A. fumigatus. Ao longo de um ano, 11 pulverizações e recuperações mensais dos 4 isolados a partir de amostras de solo permitiram monitorar possível emergência de resistência aos triazólicos: difenoconazol, metconazol, tebuconazol, itraconazol, voriconazol e posaconazol. A busca da resistência foi feita pela determinação de MIC, tanto de fungicidas quanto de fármacos triazólicos e pesquisa de mutações gênicas no gene cyp51A e sua região promotora. O gene cyp51A sequenciado para um desses isolados não apresentou mutações. Conclui-se que a pulverização de difenoconazol não resultou em alteração de MIC de triazóis nas cepas expostas indicando que não houve seleção de fenótipos resistentes. Outra possibilidade é que, de fato, o tempo de exposição, ou volume/concentração de fungicida não foram suficientes para pressão seletiva. (AU)

Clinical resistance to triazoles, drugs used for invasive aspergillosis, is a growing problem and its relationship with environmental issues is under discussion. The triazolics are also used to control phytopathogenic fungi and the same problem of increasing resistance is present. The relationship between resistance in clinical and environmental strains is based on the same mode of action of the compounds, so that exposure to one triazole compound may select another resistant strain, characterizing "crossresistance". In environmental strains of the main etiological agent of aspergillosis-Aspergillus fumigatus, classified as azole resistant, mutations associated with the cyp51A gene have been described. This gene encodes the key enzyme in the production of cell membrane ergosterol-14-demethylase, target of all azole compounds. Mutations in the cyp51A gene result in enzyme production with changes in its three-dimensional conformation leading to lower target azole affinity and antifungal resistance. The induction of such mutations was attributed to the extensive exposure of environmental strains to triazole fungicide pesticides. Studies in several countries suggest that these products exert selective pressure on resistant A. fumigatus phenotypes. The objective of this study was to evaluate the hypothesis of the environmental origin of triazole resistance mechanisms in 2 clinical isolates and 2 environmental isolates of A. fumigatus, modulated by exposure to an agricultural fungicide. Experimental garden model, organic part and part exposed to triazolic pesticide, was built and inoculated with the 4 isolates, all non-mutant wild A. fumigatus. Over a year, 11 sprayings and monthly recoveries of the 4 isolates from soil samples allowed monitoring of possible emergence of resistance to triazoles: difenoconazole, metconazole, tebuconazole, itraconazole, voriconazole and posaconazole. The search for resistance was made by the search for gene mutations that, so far, has not been found. MIC determination of both fungicides and triazole drugs against isolates, exposed and recovered by the 5th month. The cyp51A gene sequenced for one of these isolates showed no mutations. By microsatellite technique, the same isolate was not similar to the seeded isolate. It was concluded that difenoconazole spraying did not result in alteration of triazole MIC in the exposed strains indicating that there was no selection of resistant phenotypes. On the other hand, it was not possible to correlate the inoculated strain with the recovered strain and evaluated for MIC. Therefore, other recovered strains should be compared to inoculated strains for more grounded conclusion. Another possibility is that, in fact, exposure time, or fungicide volume / concentration was not sufficient for selective pressure. (AU)

Elastin increases biofilm and extracellular matrix production of Aspergillus fumigatus

Abstract Aspergillus fumigatus is an opportunistic saprobe fungus that accounts for 90% of cases of pulmonary aspergillosis in immunosuppressed patients and is known for its angiotropism. When it reaches the respiratory tract, A. fumigatus interacts with structural components and blood vessels of the lungs, such as elastin. To understand the effect of this structural component, we examined the effect of elastin on the production and development of the biofilm of A. fumigatus. In RPMI containing 10 mg/mL of elastin, a significant increase (absorbance p < 0.0001; dry weight p < 0.0001) in the production of biofilm was observed in comparison to when RPMI was used alone, reaching a maximum growth of 18.8 mg (dry weight) of biofilm in 72 h. In addition, elastin stimulates the production (p = 0.0042) of extracellular matrix (ECM) and decreases (p = 0.005) the hydrophobicity during the development of the biofilm. These results suggest that elastin plays an important role in the growth of A. fumigatus and that it participates in the formation of thick biofilm.

Cloning and expression of an endoglucanase gene from the thermotolerant fungus Aspergillus fumigatus DBiNU-1 in Kluyveromyces lactis

Abstract An intronless endoglucanase from thermotolerant Aspergillus fumigatus DBINU-1 was cloned, characterized and expressed in the yeast Kluyveromyces lactis. The full-length open reading frame of the endoglucanase gene from A. fumigatus DBiNU-1, designated Cel7, was 1383 nucleotides in length and encoded a protein of 460 amino acid residues. The predicted molecular weight and the isoelectric point of the A. fumigatus Cel7 gene product were 48.19 kDa and 5.03, respectively. A catalytic domain in the N-terminal region and a fungal type cellulose-binding domain/module in the C-terminal region were detected in the predicted polypeptide sequences. Furthermore, a signal peptide with 20 amino acid residues at the N-terminus was also detected in the deduced amino acid sequences of the endoglucanase from A. fumigatus DBiNU-1. The endoglucanase from A. fumigatus DBiNU-1 was successfully expressed in K. lactis, and the purified recombinant enzyme exhibited its maximum activity at pH 5.0 and 60 °C. The enzyme was very stable in a pH range from 4.0 to 8.0 and a temperature range from 30 to 60 °C. These features make it suitable for application in the paper, biofuel, and other chemical production industries that use cellulosic materials.

Differential role of gpaB and sidA gene expressions in relation to virulence in Aspergillus species from patients with invasive aspergillosis

Abstract The virulence genes in invasive aspergillosis (IA) have not been analyzed adequately. The present study was designed to evaluate the expression of gpaB and sidA genes, which are important virulence genes in Aspergillus spp. from bronchoalveolar lavage (BAL) samples. Direct examination and culture on Czapek Agar and Sabouraud Dextrose Agar media were performed for 600 BAL specimens isolated from patients with possible aspergillosis. A Galactomannan ELISA assay was also carried out. The expression levels of the gpaB and sidA genes in isolates were analyzed using quantitative real-time PCR (qRT-PCR). We identified 2 species, including Aspergillus flavus (A. flavus) and Aspergillus fumigatus (A. fumigatus) in 25 positive samples for invasive aspergillosis as validated using GM-ELISA. A. flavus is the main pathogen threatening transplant recipients and cancer patients worldwide. In this study, A. flavus had low levels of the gpaB gene expression compared to A. fumigatus (p = 0.006). The highest sidA expression was detected in transplant recipients (p = 0.05). There was no significant correlation between sidA expression and underlying disease (p = 0.15). The sidA and gpaB gene expression patterns may provide evidence that these virulence genes play important roles in the pathogenicity of Aspergillus isolates; however, there are several regulatory genes responsible for the unexpressed sidA and gpaB genes in the isolates.

Exercício físico ou atividade física: qual apresenta maior associação com a percepção da qualidade do sono de adolescentes? / Exercise or physical activity: which is more strongly associated with the perception of sleep quality by adolescents?

RESUMO Objetivo: Analisar a associação do exercício físico e da atividade física com a percepção da qualidade do sono em adolescentes. Métodos: Trata-se de um estudo com abordagem quantitativa que integra o levantamento epidemiológico transversal de base escolar e abrangência estadual cuja amostra foi constituída por 6.261 adolescentes (14 a 19 anos), selecionados por meio de uma estratégia de amostragem aleatória de conglomerados. Os dados foram coletados a partir do questionário Global School-based Student Health Survey. O teste do qui-quadrado e a regressão logística binária foram utilizados nas análises dos dados. Resultados: Na amostra, 29% dos adolescentes não faziam exercício e não foram classificados como fisicamente ativos. Os adolescentes que não praticavam exercício físico tinham mais chances de apresentar uma percepção negativa da qualidade do sono (OR 1,13, IC95% 1,04-1,28; p=0,043). Não foi encontrada associação entre o nível de atividade física e a percepção da qualidade do sono (OR 1,01, IC95% 0,89-1,14; p=0,868). Ao serem avaliadas as práticas de forma isolada ou simultânea, constatou-se que aqueles que praticavam exercício físico apresentavam menor chance de terem uma percepção negativa da qualidade do sono (OR 0,82, IC95% 0,71-0,95) e, ao praticarem exercício e, paralelamente, terem uma vida fisicamente ativa, essas chances diminuíam ainda mais (OR 0,79, IC95% 0,68-0,93). Conclusões: Ser classificado como fisicamente ativo, por si só, não foi suficiente para uma melhor percepção da qualidade do sono, pois apenas a prática de exercício físico apresentou tal associação.

ABSTRACT Objective: To analyze the association of exercises and physical activity with the perception of sleep quality by adolescents. Methods: This is a cross-sectional epidemiological survey with statewide coverage, whose sample was composed of 6,261 adolescents (14-19 years old) who were selected by random sampling of conglomerates. The Global School-Based Student Health Survey questionnaire was used for data collection. The chi-square test and the binary logistic regression were applied for data analyses. Results: In the sample, 29% of adolescents did not exercise and were not classified as physically active. Adolescents who did not exercise were more likely to present a negative perception of sleep quality (OR 1.13, 95%CI 1.04-1.28, p=0.043). No association between the level of physical activity and the perception of sleep quality was found (OR 1.01, 95%CI 0.89-1.14, p=0.868). Those who practiced exercises only had less chance of perceiving sleep quality as poor (OR 0.82, 95%CI 0.71-0.95). However, those who practiced exercise and had a physically active life had less chances of having a negative perception of their sleep (OR 0.79, 95%CI 0.68-0.93). Conclusions: Practicing physical activity alone was not enough to increase the chances of positive sleep quality perception. Only physical exercise had a positive association with sleep quality perception.

Can Aspergillus fumigatus conidia cause false-positive results in the galactomannan enzyme immunoassay test?

Abstract INTRODUCTION: Several factors can cause false-positive results in the galactomannan (GM) test; however, others remain unknown. Presently, the impact of airborne contamination by Aspergillus conidia during enzyme-linked immunosorbent assay (ELISA) remains uninvestigated. METHODS: We studied 12 A. fumigatus isolates. Fungal conidia were serially diluted and tested for GM detection using the Platelia® Aspergillus enzyme immunoassay (EIA). RESULTS: The conidia concentration required for an EIA-positive result was 4.8 × 103 (median). CONCLUSIONS: This is the first study to evaluate the impact of environmental contamination on the Platelia® Aspergillus EIA assay. Only massive contamination can interfere with GM optical readings, suggesting that environmental contamination does not cause false-positive test results.

Estudio clínico y micológico de otomicosis en diabéticos: una serie de 17 casos / Clinical and mycological study of otomycosis in diabetics: series of 17 cases

RESUMEN Introducción: La otomicosis en pacientes inmunosuprimidos esta caracterizada por ser bilateral y ser causada por candida. Pocos estudios comparan las características micológicas encontradas en la microscopía directa y el cultivo. Objetivo: Identificar las características clínicas y micológicas de la otomicosis en pacientes diabéticos. Material y método: Estudio transversal en centro hospitalario de segundo nivel. Criterios de inclusión: pacientes diabéticos con diagnóstico clínico de otomicosis. Intervención: la muestra se examinó directamente bajo el microscopio y se cultivó. Resultados: Se incluyeron 17 pacientes, 10 mujeres y 7 hombres con una edad media de 47,5 años. Los síntomas predominantes fueron hipoacusia en 91,4% (n =16), prurito en 82,4% (n =14), otorrea en 76,5% (n=13)y otalgia en 70,6% (n =12). Afección bilateral se encontró en 47,1% (n =8). Estudio directo al microscopio mostró levaduras en 94,1% (n =16) y 5,9% mostró aspergillus (n =1). Cándida fue el género más comúnmente encontrado en los cultivos y en el examen directo microscópico con 94,1% (n =16) y Candida albicans la especie más común con 88,2% (n =15). Conclusión: Candida albicans es el agente etiológico más común en pacientes diabéticos con otomicosis. Su presentación clínica más frecuente es hipoacusia, prurito y otorrea. El examen directo identificó adecuadamente a los géneros fúngicos.

ABSTRACT Introduction: Otomycosis in immunocompromised patients is characterize by its bilateral course and the predominant etiologic agent is Candida. Few studies compare the mycological features between microscopic direct exam and culture. Aim: To identify the clinical and mycological characteristics of otomycosis in diabetic patients. Material and method: Transversal study. Secondary care center. Inclusion criteria: diabetic patients with clinical diagnosis of otomycosis. Intervention: Direct examination under a microscope of the ear sample and culture. Results: We included 17 patients, 10 women, 7 men with a mean age of 47.5 years. Symptoms were hearing loss 94.1% (n = 16), pruritus 82.4% (n =14) otorrhoea 76.5% (n =13) and otalgia 70.6% (n =12). Bilateral involvement was found in 47.1% (n =8). Direct microscopic study found 94.1% of yeast (n =16) and 5.9% of Aspergillus (n =1). Candida was the most common fungal genus in culture and microscopic exam with 94.1% (n =16) of cases and Candida albicans was the most common species in 88.2% (n =15) cases. Conclusion: Candida albicans is the most common etiologic agent in diabetic patients with otomycosis. Main symptoms were hearing loss, itching and otorrhea. Direct exam correctly identified the fungal genus.

In vitro antifungal susceptibility of clinical and environmental isolates of Aspergillus fumigatus and Aspergillus flavus in Brazil

ABSTRACT The in vitro susceptibility of 105 clinical and environmental strains of Aspergillus fumigatus and Aspergillus flavus to antifungal drugs, such as amphotericin B, azoles, and echinocandins was evaluated by the broth microdilution method proposed by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Following the EUCAST-proposed breakpoints, 20% and 25% of the clinical and environmental isolates of A. fumigatus, respectively, were found to be resistant to itraconazole (Minimal Inhibitory Concentration, MIC > 2.0 mg/L). Voriconazole showed good activity against A. fumigatus and A. flavus strains, except for one clinical strain of A. fumigatus whose MIC was 4.0 mg/L. Posaconazole (≤0.25 mg/L) also showed appreciable activity against both species of Aspergillus, except for six A. fumigatus strains with relatively higher MICs (0.5 mg/L). The MICs for Amphotericin B ranged from 0.06 to 1.0 mg/L for A. fumigatus, but were much higher (0.5-8.0 mg/L) for A. flavus. Among the echinocandins, caspofungin showed a geometric mean of 0.078 and 0.113 against the clinical and environmental strains of A. flavus, respectively, but had elevated minimal effective concentrations (MECs) for seven of the A. fumigatus strains. Anidulafungin and micafungin exhibited considerable activity against both A. fumigatus and A. flavus isolates, except for one environmental isolate of A. fumigatus that showed an MEC of 1 mg/L to micafungin. Our study proposes that a detailed investigation of the antifungal susceptibility of the genus Aspergillus from different regions of Brazil is necessary for establishing a response profile against the different classes of antifungal agents used in the treatment of aspergillosis.