Significance of Aspergillus spp. isolation in defining cases of COVID-19 Associated Pulmonary Aspergillosis - CAPA

ABSTRACT COVID-19-Associated Pulmonary Aspergillosis (CAPA) is a relatively common complication in patients with severe forms of the disease caused by the SARS-CoV-2 virus. Diagnosing and confirming CAPA is challenging. In this study, Aspergillus spp. isolation in respiratory specimens from patients with COVID-19 was evaluated for identifying cases of CAPA. In 2020-2021, 17 Aspergillus spp. were isolated from 15 COVID-19 patients admitted to a university hospital in Brazil. Patient records were retrospectively reviewed to obtain clinical-epidemiological data and other markers of Aspergillus spp. infection and then compared with the ECMM/ISHAM criteria for defining CAPA. Probable CAPA was defined in 5/10 patients, who had Aspergillus spp. isolated from Bronchoalveolar Lavage (BAL) or a positive galactomannan blood test. Additionally, anti-Aspergillus antibodies were detected in two of these patients, during active or follow-up phases of CAPA. In another seven patients with Aspergillus spp. isolated from tracheobronchial aspirate or sputum, CAPA was presumed, mainly due to deterioration of clinical conditions and new lung imaging suggestive of fungal infection. Antifungal agents to control CAPA, particularly voriconazole, were used in 9/15 cases. In cases of probable CAPA and remaining patients, clinical conditions and comorbidities were similar, with lethality being high, at 60% and 71%, respectively. The number of CAPA cases defined by scientific criteria was lower than that assumed in the clinical context. This was largely due to the lack of BAL collection for fungal culture and the non-intensive use of other markers of invasive aspergillosis. The isolation of Aspergillus spp. in different respiratory specimens should alert clinicians to the diagnosis of CAPA.

Micosis en tiempos de COVID-19 / Mycosis in times of COVID-19

El brote mundial del SARS-CoV-2, descrito a partir del 2019, provocó la pandemia de COVID-19, originando un riesgo para la salud de las personas, una amenaza a la vida y una emergencia de salud pública internacional, que hasta Julio del 2021 no se ha logrado controlar. La coinfección en estos pacientes, por virus, bacterias y hongos, aumenta la dificultad de diagnóstico, tratamiento y pronóstico de la enfermedad. Es importante profundizar los conocimientos sobre el virus SARS-CoV-2 y las coinfecciones que podrían presentarse, en particular, en pacientes con COVID-19 que presentan micosis. El objetivo de esta revisión bibliográfica es, determinar la importancia de las micosis, como enfermedad oportunista, en pacientes con COVID-19. Se realizó una revisión bibliográfica sistemática, en la base de datos "PubMed-NCBI". Se utilizaron las palabras claves: "COVID-19", "SARS-CoV-2", "Coronavirus", "COVID-19 and coinfection", "Mycosis", "Aspergillus spp.", "Candida spp.", "COVID-19 and Aspergillus spp.", "COVID-19 and Candida spp.". Del análisis de la bibliografía, se concluye la importancia de las micosis respiratorias, originadas por diversos hongos en pacientes con COVID-19. Hay poca información del manejo de estas, siendo necesario fortalecer la investigación de la coinfección, para así, mejorar los flujogramas de sospecha clínica, contribuyendo a diagnósticos, tratamientos precisos y fomentar la prevención frente a esta pandemia.

The global outbreak of SARS-CoV-2, described as of 2019 whose expansion caused the COVID-19 pandemic, has created a risk to people's health, presenting itself as a threat to life and an international public health emergency, which to the date cannot be controlled. Coinfection in these patients, by viruses, bacteria and fungi, increases the difficulty of diagnosis, treatment and prognosis of the disease. It's important to deepen the knowledge about the SARS-CoV-2 virus and the co-infections that could occur, in particular, in patients with COVID-19 who present with mycosis. The objective of this bibliographic review is to determine the importance of mycosis, as an opportunistic disease, in patients affected by SARS-CoV-2. A systematic bibliographic review was carried out in the "PubMed-NCBI" database, using the keywords and / or headings: "COVID-19", "SARS-CoV-2", "Coronavirus", "COVID-19 and coinfection" ,"Mycosis", "Aspergillus spp.", "Candida spp." COVID-19 and Aspergillus spp.", "COVID-19 and Candida spp.". From the analysis of the literature, one can conclude the importance of respiratory mycoses, caused by various fungal pathogens in patients with COVID-19. The disease was described in 2019 and there is few a information on cases and their management, making it necessary to strengthen the investigation of coinfection in these patients, in order to improve the flow charts of clinical suspicion, contributing to diagnoses, accurate treatments and promoting prevention against to this pandemic.

Microbiological And Epidemiological Profile Of Keratomycosis In A Tertiary Care Hospital – A Prospective Analysis

Introduction: Microbial keratitis is predominantly an opportunistic, serious ocular infectious disease that can lead to significant vision loss and ophthalmic morbidity.1 The fungi are significant pathogens causing ocular infections due to their frequent involvement and difficulty in establishing definitive diagnosis.. Material and Methods: We prospectively analyzed 106 cases of keratomycosis in our tertiary care hospital. Corneal scrapings were collected, processed and fungal pathogens were identified by standard laboratory techniques. Results: Out of 106 suspected cases, culture was positive in 31 cases. Aspergillus species were the most frequent isolates (51.61%). Next to this were Candida spp. (19.35%) followed by Fusarium spp., Alternaria spp., Mucor spp , Penicillium spp, Drechslera spp and Exserohilum spp . Males were more affected than females and trauma was the most common predisposing factor. Conclusions: This study highlights important risk factors and organisms responsible for mycotic keratitis.

Current status of invasive fungal infection in pediatric intensive care unit / 中国小儿急救医学

The invasive fungal infection( IFI)in PICU has increased steadily during the recent years. Candida spp. and Aspergillus spp. are the most frequently fungi in children. Candida spp. is the leading cause and invasive Candida spp. Infection( ICI)is approximately five times frequency than invasive Aspergillus spp. Infection( IAI). The attributable mortality of ICI or IAI remains different mainly because of different basic diseases. Stay in the PICU presents risk factors for ICI especialy using central venous catheter,parenteral nutrition,dialysis,mechanical ventilation,and prolonged antibiotics application. The patients with hematologic malignancies and leukemia are higher prevalence of IAI who were treated with cytotoxic or immunosuppres-sive drugs,broad-spectrum antibiotics and stem cell transplantation. The most important challenge remains to propose targeted prophylaxis and to identify IFI earily in high risk critically ill children in PICU.

Serological monitoring of antibodies for an early diagnosis ofaspergillosis in captive penguins / Monitoramento sorológico com detecção de anticorpos para diagnóstico precoce da aspergilose em pinguins em cativeiro

This study aimed to evaluate the efficacy of detection of anti-Aspergillus fumigatus antibodies in captive penguins by double radial agar gel immunodiffusion (AGID) for the aspergillosis diagnosis. We included 134 Magellanic penguins (Spheniscus magellanicus) in rehabilitation at the Center for Recovery of Marine Animals (CRAM / FURG). All of them were monitored by AGID weekly until its final destination (death or release), totalizing 660 serum samples studied. All animals were clinically accompanied and post-mortem examinations was performed in penguins that died during the studied period. A total of 28% (37/134) of the penguins died, 89.2% (33/37) due to aspergillosis, 11% (4/37) by other causes and 97 were released. From the 33 animals with proven aspergillosis, 21 presented anti- A. fumigatus antibodies by AGID, being the average interval between death and positive AGID 16.4 days. Twelve animals with negative serology died of aspergillosis. The sensitivity and specificity rates were 63.6% and 95% respectively, and the positive and negative predictive values were 80.7% and 88.9% respectively. These data demonstrate that the serological monitoring for detection of antibodies by AGID can be an important tool for the diagnosis of aspergillosis in penguins.

Este estudo teve como objetivo avaliar a eficácia da detecção de anticorpos anti- Aspergillus fumigatus em pinguins em cativeiro por imunodifusão radial dupla em gel de ágar (IDGA) para diagnóstico da aspergilose. Foram incluídos 134 pingüins de Magalhães (Spheniscus magellanicus) em reabilitação no Centro de Recuperação de Animais Marinhos (CRAM/FURG), que foram monitoradas por IDGA, semanalmente, até o seu destino final (morte ou de liberação), totalizando 660 amostras de soro estudadas. Todos os animais foram acompanhados clinicamente e exames post mortem foram realizados em pingüins que vieram a óbito durante o período de estudo. Um total de 28% (37/134) dos pinguins foram a óbito, 89,2% (33/37) de aspergilose, 11% (4/37) de outras causas, e 97 foram liberados. A partir dos 33 animais com aspergilose comprovada, 21 apresentaram anticorpos anti- A. fumigatus por IDGA, sendo o intervalo médio entre a morte e IDGA positivas 16,4 dias. Doze animais com sorologia negativa vieram a óbito por aspergilose. As taxas de sensibilidade e especificidade foram de 63,6% e 95%, respectivamente, e os valores preditivos positivos e negativos foram de 80,7% e 88,9 %, respectivamente. Estes dados demonstram que o monitoramento sorológico para detecção de anticorpos por IDGA pode ser uma ferramenta importante no diagnóstico de aspergilose em pinguins.

Incidência de aflatoxinas no amendoim (Arachis hypogaea L) cru em casca da região da Alta Paulista-SP, durante o período de 2011 a 2012 / Incidence of aflatoxins in the raw peanut (Arachis hypogaea L) in shell of Alta Paulista-SP region during the period from 2011 to 2012

As aflatoxinas são metabólitos secundários produzidos pelas espécies do gênero Aspergillus e a frequente ingestão de alimentos contendo teores de aflatoxinas tem sido correlacionada às neoplasiashepáticas pela International Agency for Reseach on Cancer (IARC). Foram analisadas 966 amostras de amendoim cru em casca, coletadas no período de fevereiro a maio, correspondente à época de colheita das safras 2010-2011 e 2011-2012. Para detecção de aflatoxinas foram utilizadas as metodologias de coluna de imunoafinidade acoplada à fluorometria (CIA) e cromatografia líquida de alta eficiência(CLAE), seguindo-se os parâmetros estabelecidos pela Agência Nacional de Vigilância Sanitária. Foram detectadas aflatoxinas acima do limite máximo tolerado pela legislação em 12,6 % e 19,5 % das amostras,respectivamente nas safras de 2011 e 2012. Na safra de 2010-2011, a faixa de contaminação foi de 21-52 mg/kg, e na safra de 2011-2012 a faixa de contaminação variou de 28-260 mg/kg, cujo valor é cinco vezes maior quando comparado ao obtido em 2011. Os maiores índices de contaminação foram encontrados nos períodos com menor índice pluviométrico, tanto em 2011 quanto em 2012. Por conseguinte, a realização de rastreamento é necessária durante toda a cadeia produtiva do amendoim, com a finalidade de propiciar alimentos seguros à saúde humana e animal...

Aspergilosis diseminada: hallazgos postmortem en un caso de sepsis materna. ¿Es el embarazo un estado de inmunosupresión? / Disseminated aspergillosis: postmortem findings in a case of maternal sepsis

Se comunica un caso de muerte materno-fetal con diagnóstico de aspergilosis diseminada concluido a través del estudio de autopsia. Caso: Mujer de 20 años de edad con antecedente de preeclampsia quien en su segundo embarazo inició con dolor obstétrico asociado a fiebre y ataque al estado general. El trabajo de parto concluyó con la expulsión de un feto óbito macerado y placenta fétida, hipotonía uterina e inestabilidad hemodinámica. Fue referida a segundo nivel, cursaba con fiebre persistente, foco séptico pélvico, trastornos de la ventilación, evidencia de lesiones encefálicas por tomografía, sangrado de tubo digestivo, oliguria, datos de respuesta inflamatoria sistémica y necrosis bilateral de miembros inferiores. Falleció a los 10 días de haber sido internada. Resultados: En el examen necrológico, la División de Anatomía Patológica halló pulmones con áreas extensas de consolidación y hemorragia, corazón con vegetaciones valvulares y murales, así como cerebro con múltiples lesiones similares a infarto. La microscopía de luz reveló la presencia de hifas septadas ramificadas en ángulos de 45° que transgredían el lumen de arterias en pulmón, asentadas sobre la superficie endocárdica e invadían el miocardio, el espacio subaracnoideo y neurópilo. Dichas hifas también se identificaron en los cortes histológicos de tiroides, tráquea, estómago y riñón. Conclusión: La aspergilosis diseminada es una entidad de elevada letalidad que usualmente afecta a pacientes inmunocomprometidos, específicamente a quienes cursan con neutropenia. Si bien durante el embarazo el sistema inmune experimenta determinadas adaptaciones fisiológicas, éstas no predisponen per se al desarrollo de infecciones oportunistas.

We describe autopsy findings in a case of maternal death caused by invasive aspergillosis in its disseminated form. Case: 20 year old female with previous medical history of preeclampsia who started with malaise, pelvic pain and fever during her second pregnancy. A fetal obitus was obtained after labor although she remained with uterine hypotony and hemodynamic unsteadiness. She was referred to a general hospital subsisting with fever, acute pelvic sepsis, deteriorated ventilatory function, cerebral lesions visualized by tomography, gastrointestinal bleeding, oliguria and systemic inflammatory response with bilateral acral lower extremities necrosis. She died ten days after her arrival and autopsy was authorized. Results: Necropsy uncovered lungs with consolidated areas and extensive haemorrhage, valvular and mural heart vegetations and numerous cerebral lesions with infarct-like appeareance. Light microscopy revealed the presence of septate hyphae with regular acute angle branching invading lung microvasculature, endocardial surface, myocardium, Virchow-Robin space and neuropil. The hyphae were also identified in thyroid, trachea, gastric mucosa and kidney histological sections. Conclusion: Disseminated aspergillosis is a high-mortality infectious process that usually affects immunocompromised patients, specially those coursing with neutropenia. Despite immunological changes experienced as part of normal pregnancy, these per se do not predispose the mother to infections caused by opportunistic pathogens.

Antifungal susceptibility testing of few medicinal plant extracts against Aspergillus spp. and Microsporum sp.

Mycoses are fungal infections, the incidence of which in immunocompromised patients is currently devastating and the drugs available at hand are reported to exhibit side effects. To surmount the prevailing difficulty of complete eradication of the fungal infection, exploring new arena is a requisite and nature’s wealth can be one area that may lead to a cure for fungal infections. In the present study, the whole methanolic extracts of few medicinal plants were evaluated against Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger and the dermatophyte Microsporum gypseum isolated from the soil. Enicostemma littorale and Wrightia tinctoria exhibited significant (P<0.001) inhibition of about 48% against A. niger. Eupatorium odoratum inhibited both A. fumigatus (52%) and A.flavus (32%) whereas Enicostemma littorale showed about 54% of significant (P<0.05) inhibition against A. fumigatus. Significant inhibition of M. gypseum was exhibited only by Sphaeranthus indicus flower extracts (65% at P<0.05) which was even higher than the inhibition exhibited by positive control ketaconazole (49%) at 0.1 mg/ml concentration. This study demonstrates that among the medicinal plants evaluated E.littorale, W.tinctoria, E.odoratum and S.indicus flower exhibited significant antifungal activity against the tested organisms.

Evaluación de las enzimas celulolíticas producidas por hongosnativos mediante fermentación en estado sólido (SSF) utilizando residuos de cosecha de caña de azúcar / Evaluation of cellulolytic enzymes produced by native fungi through solid state fermentation (SSF) using sugarcane harvesting residues

Los residuos agrícolas de cosecha de caña de azúcar (RAC), se constituyen en una materia prima alternativa para la producción de etanol carburante, dado su contenido de celulosa próximo al 40%. El aprovechamiento de la celulosa depende de la aplicación de tratamientos fisicoquímicos o bioquímicos, que permitan la liberación de la glucosa y su posterior utilización en procesos fermentativos. La hidrólisis enzimática de estos residuos requiere un complejo celulolítico producido por microorganismos, comprendido por tres actividades enzimáticas: Endoglucanasas, Exoglucanasas y β-Glucosidasas. En el presente estudio, se evaluaron las enzimas celulolíticas producidas por dos hongos nativos del género Aspergillus spp., CH 2016 y CH 2001, mediante procesos de fermentación en estado sólido utilizando como sustrato RAC pre-tratados con organosolventes (deslignificado) y sin este pre-tratamiento. La cepa CH 2016 presentó la mayor actividad endoglucanasa 11,0773 U/mL en el sustrato sin pre-tratar el día siete de fermentación; esta misma cepa, en el sustrato deslignificado presentó la mayor actividad exoglucanasa (0,042 U/mL) y celulasa total (0,287 UPF/mL) en el día cinco de fermentación. La cepa CH 2001 presentó la mayor actividad β-glucosidasa (0,1778 U/mL) en el sustrato sin pre-tratar el día cinco de fermentación. Se observó que las variables sustrato y tiempo de fermentación, inciden en la expresión de las enzimas celulolíticas obteniendo en este trabajo extractos enzimáticos que pueden llevar a cabo una acción hidrolítica sinérgica sobre la celulosa.

Sugarcane harvesting residues are considered as a raw material for fuel ethanol production due its high content of cellulose, around 40% DS. The use of cellulose depends of the application of physicochemical or biochemical treatments that allow the release of glucose and its subsequent uses in fermentation processes. The enzymatic hydrolysis of these residues requires a cellulolytic complex produced by microorganisms, including three enzymatic activities: Endoglucanases, β-Glucosidases and Exoglucanases. In the present study, cellulolytic enzymes produced by two native fungi Aspergillus spp., CH 2016 and CH 2001 was assessment, through of solid-state fermentation processes using as raw substrate RAC and pre-treated with organosolvents (delignified). Strain CH 2016 had the highest endoglucanase activity 11.0773 U/mL in the raw substrate on day seven of fermentation, the same strain, in the delignified substrate showed the highest activity exoglucanasa (0.042 U/mL ) and total cellulase (0.287 UPF/mL) on day five of fermentation. Strain CH 2001 got the highest β-glucosidase activity (0.1778 U/mL) in the substrate without pre-treatment on day 5 of fermentation. It was observed that the variables as substrate and fermentation time affected the expression of cellulolytic enzymes.

Qualidade microbiológica de castanhas de caju (Anacardium occidentale, L) industrializadas e processadas artesanalmente / Study on the microbiological quality of industrialized and handmade cashew nuts (Anacardium occidentale L)

O objetivo deste estudo foi avaliar a qualidade microbiológica de castanhas industrializadas e das castanhas artesanalmente processadas, comercializadas por ambulantes em Teresina (PI). Foram coletadas 40 amostras de castanhas, sendo 21 amostras de castanhas industrializadas de três marcas (“A”, “B” e “C”) e 19 amostras de castanhas processadas artesanalmente (“D”), nas quais foram realizadas a determinação de coliformes a 35 °C e a 45 °C (NMP/g), a pesquisa de Salmonella spp. e a contagem de fungos. As amostras da marca “D” apresentaram maiores valores de coliformes a 35 °C (1,16 × 101 NMP/g); para coliformes a 45 °C, foram detectados valores de 7,0 NMP/g, e de 1,22 × 102 UFC/g para fungos e leveduras. Nas amostras da marca “A”, os valores para coliformes a 35 °C e 45 °C foram de 4,0 NMP/g e, para fungos e leveduras,de 1,0 × 102 UFC/g. Foram isoladas 43 cepas fúngicas. Do gênero Aspergillus, houve maior prevalência da espécie Aspergillus niger agregados (64,7%), e as espécies P. corylophillum (33,3%) e P. citrinum (29,2%) do gênero Penicillium. As amostras de castanhas industrializadas e processadas artesanalmente apresentaram condições higiênico-sanitárias satisfatórias e de acordo com a legislação vigente.

Identification of fungi of the genus Aspergillus section nigri using polyphasic taxonomy

In spite of the taxonomy of the Aspergillus species of the Nigri Section being regarded as troublesome, a number of methods have been proposed to aid in the classification of this Section. This work aimed to distinguish Aspergillus species of the Nigri Section from foods, grains and caves on the basis in Polyphasic Taxonomy by utilizing morphologic and physiologic characters, and sequencing of ß-tubulin and calmodulin genes. The morphologic identification proved useful for some species, such as A. carbonarius and Aspergillus sp UFLA DCA 01, despite not having been totally effective in elucidating species related to A. niger. The isolation of the species of the Nigri Section on Creatine Sucrose Agar (CREA) enabled to distinguish the Aspergillus sp species, which was characterized by the lack of sporulation and by the production of sclerotia. Scanning Electron microscopy (SEM) allowed distinguishing the species into two distinct groups. The production of Ochratoxin A (OTA) was only found in the A. carbonarius and A. niger species. The sequencing of β-tubulin gene was efficient in differing most of the Aspergillus species from the Nigri Section with the exception of Aspergillus UFLA DCA 01, which could not be distinguished from A. costaricaensis. This species is morphologically similar to A. costaricaencis for its low sporulation capacity and high sclerotia production, but it differs morphologically from A. costaricaensis for its conidial ornamentation and size of vesicles. Equally, based on partial calmodulin gene sequence data Aspergillus UFLA DCA 01 differs from A. costaricaensis.

Estandarización del inóculo por desintometría para las pruebas de susceptibilidad antifungica en aspergillus spp / Standardization of inoculum by densitometry for antifungal susceptibility testing in aspergillus spp

En los últimos años, la enfermedad invasora causada por Aspergillus spp. ha constituido un problema creciente en pacientes inmunosuprimidos, debido a su elevada incidencia y mortalidad. La aparición de nuevas alternativas terapéuticas como el voriconazol y la caspofungina, aunada a la resistencia documentada de especies de Aspergillus a la anfotericina B e itraconazol, conllevan a la realización de pruebas de susceptibilidad antifúngica de aislados clínicos de Aspergillus spp. Actualmente existen dos documentos de referencia aprobados par a las pruebas de susceptibilidad de Aspergillus spp., el M38-A2 y el E.DEF 9.1. Una de las variables que influye significativamente en éstas pruebas, es la preparación del inoculo; en tal sentido, los documentos mencionados presentan diferentes metodologías, el M38-A2 refiere que la preparación del inóculo se realice mediante el uso de, mientras que E.DEF 9. 1 indica que dicha preparación se realice mediante el contaje de conidias en una cámara de Neubauer. Por otra parte, algunos investigadores han obtenido resultados controversiales al evaluar y/o comparar la reproducibilidad de las metodologías de preparación del inóculo de ambos métodos de referencia En ésta investigación se estandarizó una metodología alternativa para la preparación del inóculo, la densitometría, que resultó ser un método adecuado, sencillo y rápido. Para la preparación de los inóculos y se estableció los siguientes rangos de unidades: MacFarland: de 0.4 a 0.6 para A.flavus, y A. nidulans. De 0.3 a 0.4 para A.fumigatus, y A.terreus y de 0.2 a 0.8 para A.niger.

In recent years, invasive disease caused by Aspergillus spp. has become an increasing problem in immunosuppressed patients because of its high incidence and mortality. The emergence of new therapies such as voriconazole and caspofungin, coupled with documented resistance of Aspergillus species to amphotericin B and itraconazole, leading to the realization of antifungal susceptibility testing of clinical isolates of Aspergillus spp .Currently there are two reference documents approved for susceptibility testing of Aspergillus spp., the M38-A2 and E. DEF 9.1. One of the variables that significantly influence these tests, is the preparation of inoculum; in this way, the documents before mentioned show differ entmethodologies, the M38-A2 refers to the preparation of the inoculum was performed by using spectrophotometry, while E. DEF 9.1 indicates that this preparation is executing by counting conidia in a Neubauer chamber. On the other hand, some researchers have obtained controversial results when evaluating and or compare the reproducibility of the methods of preparing the inoculum of the two methods of reference. For this study, it was standardized an alternative methodology for the preparation of inoculum, "densitometry", and this resulting an appropriate method, simple and fast. For preparation of inocula by densitometry was established the following range s of units MacFarland: 0.4 to 0.6 for A.flavus, and A. nidulans. from 0.3 to 0.4 for A.fumigatus, and A. terreus and from 0.2 to 0.8 for A.niger.

Composición química y actividad biológica de aceites esenciales de Calycolpus moritzianus recolectado en el Norte de Santander, Colombia / Chemical composition and biological activity of essential oils calycolpus moritzianus collectd in the Norte de Santander, Colombia

Introducción: actualmente, se están llevando a cabo estudios sobre la actividad biológica de AEs de plantas de la familia Myrtaceae. El objetivo de esta investigación fue la caracterización química y la evaluación de la actividad antimicótica, antitumoral y antiviral de 40 aceites extraídos de plantas de la especie Calycolpus moritzianus, comúnmente llamada “arrayán ó cínaro”, recolectadas en varias zonas del Norte de Santander, Colombia. Materiales y métodos: la actividad antimicótica se evaluó contra: Candida krusei, C. parapsilosis, Aspergillus fumigatus y A. flavus, siguiendo las técnicas estándar EUCAST y CLSI M38-A, respectivamente, para levaduras y hongos filamentosos. La citotoxicidad se evaluó en las líneas celulares tumorales (HeLa, Jurkat) y no tumorales (Vero) mediante la técnica fotocolorimétrica del MTT. La actividad antiviral in vitro, contra el virus Herpes simplex tipo HSV-1, se realizó mediante la “técnica de titulación del punto final (EPTT)”. Resultados y Discusión: los aceites X38 y X39, de muestras recolectadas en los municipios de Chinácota y Salazar, mostraron actividad contra C. krusei con valores de concentraciones inhibitorias mínimas de 500 µg/mL. Ninguno de los aceites fue citotóxico en células no tumorales Vero. Los aceites X9 y X10, procedentes de los municipios de Salazar y Pamplonita, fueron los más citotóxicos sobre células HeLa, a concentraciones de 37,8 ± 2.4 y 40,6 ± 2,6 µg/mL, respectivamente. La reducción de la carga viral en los cultivos celulares sólo fue lograda por el aceite X11. La actividad antimicótica y antitumoral de los aceites fue correlacionada con la composición de los componentes mayoritarios, evidenciándose la actividad del Terpinen-4-ol, (-) Limoneno, α –Pineno y Linalol.

Background: Essential oils of plants of the family Myrtaceae have been evaluated for different biological activities. The aim of this research was the chemical characterization and evaluation of antifungal, antiviral and cytotoxic activity of 40 essential oils extracted from Calycolpus moritzianus, commonly called “arrayán” or “cínaro”, collected in several areas of Norte de Santander, Colombia. Methods: antifungal activity was determined following the protocols AFST-EUCAST for Candida krusei and C. parasilopsis, and CLSI-M38A for Aspergillus fumigatus and A. flavus. Cytotoxicity was evaluated on tumoral (HeLa and Jurkat), and non tumoral (Vero) cell lines by the MTT photo-colorimetric technique The in vitro antiviral activity against Herpes simplex virus HSV-1 was determined by the end-point titration technique (EPTT). Results and Discussion: X38 and X39 oils of plants collected in regions of Chinácota and Salazar regions showed activity against C. krusei with minimal inhibitory concentration values of 500 µg/mL. None of the oils was cytotoxic in no-tumor (Vero) cells . Oils X9 and X10 of plants from regions of Salazar and Pamplonita were the most cytotoxic on HeLa cells at concentrations of 37,8 ± 2,4 and 40,6 ± 2,6 µg /mL, respectively. The reduction of viral load in cell cultures only was showed by X11 oil. Antifungal and antitumor activity of these oils was correlated with the composition of the major components showing activity for Terpinen-4-ol, (-) Limonene, Linalool and α-Pinene.

Characterization of Aspergillus species based on fatty acid profiles

Cellular fatty acid (FA) composition was utilized as a taxonomic tool to discriminate between different Aspergillus species. Several of the tested species had the same FA composition and different relative FA concentrations. The most important FAs were palmitic acid (C16:0), estearic acid (C18:0), oleic acid (C18:1) and linoleic acid (C18:2), which represented 95 percent of Aspergillus FAs. Multivariate data analysis demonstrated that FA analysis is a useful tool for differentiating species belonging to genus Aspergillus. All the species analyzed showed significantly FA acid profiles (p < 0.001). Furthermore, it will be possible to distinguish among Aspergillus spp. in the Flavi Section. FA composition can serve as a useful tool for the identification of filamentous fungi.

Eficácia da clorexidina-cetrimida na desinfecção ambiental contra Aspergillus spp. / Chlorexidine-cetrimide efficacy in ambient disinfection against Aspergillus spp

Avaliou-se a ação in loco da clorexidina-cetrimida no controle de Aspergillus spp., considerando-se a influência de fatores climáticos e populacionais, em um centro de recuperação de animais marinhos. Durante dois anos, realizaram-se colheitas de amostras de ar por meio da técnica de sedimentação, com implantação no segundo ano de um programa de desinfecção com clorexidina-cetrimida. Os resultados do isolamento fúngico nos dois anos foram comparados estatisticamente pelo programa Epinfo 8.0. Demonstrou-se que o isolamento de Aspergillus spp. não apresentou relação significativa com fatores climáticos e populacionais e que a diminuição da concentração de conídios de Aspergillus spp. do ambiente ocorreu devido ao programa eficaz de desinfecção com a clorexidina-cetrimida.

The study evaluated the in loco activity of chlorexidine-cetrimide for environmental control of Aspergillus spp. considering the climatic influence and population variations, in a recuperation center of marine animals. Samples of air were collected throughout two years by the sediment method, and the disinfection program was established in the second year. Data collected during the two years were statistically compared by Epinfo 8.0. program. It was demonstrated that both population and climate had no influence on the fungal isolation, and the disinfection program with chlorexidine-cetrimide was considered effective in the reduction of conidia Aspergillus concentration indoor.

Keratinolytic Activity of Five Aspergillus Species Isolated from Poultry Farming Soil in Korea

Various soil samples were collected from twenty-four areas of ten different poultry farms in Korea and screened for prevalence of keratinolytic fungi. Fourteen species of feather-associated fungi belonging to ten genera Acremonium, Alternaria, Aspergillus, Cladosporium, Curvularia, Fusarium, Monascus, Mucor, Penicillum, and Verticillium isolated from poultry soils were grown on keratin medium. Especially, Aspergillus spp. populations associated with the soil sample is 1x10(5) cfu/g. A. flavus, A. fumigatus, A. niger, A. nidulans, and A. terreus could utilize keratin of chicken feather and degrade it, producing sulphydryl-containing compounds detected as keratinase, cysteine and total proteins. Keratinolytic activities of five Aspergillus species also changed the pH of the medium more alkaline than those that were less keratinolytic.

Evaluation of Brazilian terrestrial Aspergillus strains for Mycotoxin production / Avaliação de linhagens terrestres brasileiras de Aspergillus paraprodução de micotoxinas

Screening tests for aflatoxins B1, B2, G1 and G2, ochratoxin A and sterigmatocystin production were performed in 13 strains of Aspergillus spp, isolated from the terrestrial environment in the Brazilian Atlantic Rainforest (São Paulo State/Brazil). Coconut agar medium and moistened corn were employed as substrates. The fungal extracts obtained from both media were submitted to thin-layer chromatography and the toxins were estimated according to the intensity of their fluorescence observed under UV light. None of the tested strains presented any of the mentioned mycotoxins. Because many unknown fluorescente spots were present, it was necessary to proceed a confirmation step using multiple chromatography, two dimensional chromatography and derivatization. In view of the accuracy of the employed methods and the presence of many unknown fluorescent spots, the need of further studies on the production of others mycotoxins of fungi isolated under tropical conditions is justified. (AU)

Testes de triagem para verificar a produção de aflatoxinas B1, B2, G1 e G2, ocratoxina A eesterigmatocistina foram conduzidos em 13 linhagens de Aspergillus spp, isoladas do ambiente terrestreda Mata Atlântica Brasileira (SP/Brasil). Os meios de agar côco e milho umidificado foram os substratostestados neste estudo. Os extratos dos fungos obtidos a partir dos dois substratos foram submetidos àcromatografia em camada delgada e as micotoxinas estimadas de acordo com as fluorescências apresentadassob luz ultravioleta. Nenhuma das linhagens testadas apresentou produção das micotoxinas mencionadas. Foi necessário acrescentar uma etapa de confirmação, usando múltipla cromatografia, cromatografia bidimensional e derivação. Tendo em vista a eficiência da metodologia aqui empregada e da presença de muitos pontos fluorescentes desconhecidos, justifica-se a necessidade da ampliação dos estudos sobrea produção de outras micotoxinas em fungos isolados de ambientes tropicais. (AU)

In Vitro Activities of LY303366 against Clinical Isolates of Candida spp., Cryptococcus neoformans and Aspergillus spp / 감염

BACKGROUND: Fungal infection became more prevalent with the increase of compromised patients, requiring less toxic potent antifungal agents. LY303366, a new semisynthetic antifungal agent of echinocandin class, was reported to be active against Candida spp. and filamentous fungi. METHODS: In vitro activities of LY303366 were determined against clinical isolates of fungi by NCCLS broth microdilution test using RPMI 1640 medium. RESULTS: MIC90s of LY303366 were 0.03 microgram/mL for C. albicans, 2 microgram/mL for C. parapsilosis, 0.12 microgram/mL for C. tropicalis. LY303366 was more active against C. albicans and C. tropicalis than amphotericin B, 5-fluorocytosine, or fluconazole. It was less active than other agents against C. neoformans. MIC range of LY303366 against Aspergillus spp. was 8->16 microgram/mL. CONCLUSION: Very high in vitro activity of LY303366 against recent clinical isolates of Candida spp. including fluconazole resistant ones, suggests its usefulness for the treatment of candidal infections.