How a discerning cytological examination can aid in the diagnosis of infectious diseases: case reports

Infections caused by uncommon and resistant pathogens in unusual sites have been increasingly reported in medical literature. We describe four cases of rare cytological findings and clinical impact for patients. In the first case, Aspergillus sp and Pneumocystis jirovecii were observed in the bronchoalveolar lavage of a patient with severe systemic lupus. In the second and third cases, we describe the presence of Trichomonas sp and Strongyloides sp larvae in samples of pleural and peritoneal fluid, respectively. The fourth report is about a patient with a wrist subcutaneous nodule whose synovial aspiration and cytology revealed the presence of brown septate hyphae. The early identification of the infectious agent in the cytological examination was essential for the introduction and/or re-adaptation of therapy in the four cases described. Patients in this report were immunocompromised with severe comorbidities, conditions often associated with unfavorable clinical outcomes.

Diversity and antimicrobial activity of culturable fungi associated with sea anemone Anthopleura xanthogrammica

Background: The main objective of this study was to isolate fungi associated with Anthopleura xanthogrammica and measure their antimicrobial and enzymatic activities. A total of 93 fungal strains associated with A. xanthogrammica were isolated in this study, of which 32 isolates were identified using both morphological characteristics and internal transcribed spacer (ITS) sequence analysis. The antibacterial activities of 32 fungal isolates were tested against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Edwardsiella tarda, Vibrio harveyi, Fusarium oxysporum, and Pyricularia oryzae by agar diffusion assay. Extracellular hydrolytic enzyme activities of the fungal isolates were determined by agar diffusion assays. Enzyme activities were detected from clear halo size. Results: The isolated fungi belonged to 18 genera within 7 taxonomic orders of 1 phylum. The genera Aspergillaceae were the most diverse and common. The antimicrobial activities of 32 isolates were evaluated, and 19 (59.4%) of fungi isolate displayed unique antimicrobial activities. All fungal strains displayed at least one enzyme activity. The most common enzyme activities in the fungi isolates were amylase and protease, while the least common were pectinase and xylanase. Conclusions: This is first report on the sea anemone-derived fungi with antimicrobial and enzyme activities. Results indicated that sea anemone is a hot spot of fungal diversity and a rich resource of bioactive natural products.

Confirmed Invasive Pulmonary Aspergillosis and COVID-19: the value of postmortem findings to support antemortem management

Abstract We present postmortem evidence of invasive pulmonary aspergillosis (IPA) in a patient with severe COVID-19. Autopsies of COVID-19 confirmed cases were performed. The patient died despite antimicrobials, mechanical ventilation, and vasopressor support. Histopathology and peripheral blood galactomannan antigen testing confirmed IPA. Aspergillus penicillioides infection was confirmed by nucleotide sequencing and BLAST analysis. Further reports are needed to assess the occurrence and frequency of IPA in SARS-CoV-2 infections, and how they interact clinically.

Applicability of the Platelia EIA® Aspergillus test for the diagnosis of aspergilosis in penguins / Aplicabilidade do Platelia EIA® Aspergillus como teste diagnostico da aspergilose em pinguins

Abstract Even today, an effective diagnostic test for aspergillosis in penguins is unknown, being the gold standard post-mortem examinations. The fungal antigen galactomannan (GM) has been used as a biomarker of disease in humans and is detected by the Platelia Aspergillus EIA (BioRad)®, a commercial kit based on the sandwich ELISA technique. It is standardized for use in neutropenic patients, however studies have demonstrated its usefulness also possible for birds. The aim of our study was to evaluate the effectiveness of Platelia Aspergillus EIA® test (BioRad-US) in the diagnosis of aspergillosis in Magellanic penguins, determining sensitivity, specificity, and positive and negative predictive values for different cut-off points. Were included in the study, blood serum samples (n = 29) Magellanic penguins in captivity that died by aspergillosis. Detection of GM was performed following manufacturer's instructions and the GM index was obtained by dividing the average value of OD of the duplicate of the clinical sample by duplicate OD of the average value of the cut-off sample provided by the kit. Through information database results were obtained for the presence of anti-Aspergillus fumigatus antibodies detected by agar gel immunodiffusion (AGID) for all serum samples. Results were analyzed using chi-square test and Kruskal-Wallis from SPSS 20.0, IBM®. ROC curve was obtained and from this, rates of sensitivity, specificity, positive and negative predictive values were also calculated based on four different cutoff points (0.5, 1.0, 1.5 and 2.0). The serum GM index did not differ between animals of the case and control group (pkw =0.097). In determining the ROC curve for serum GM detection the value of area under the curve was 0.635. From the values ​​determined by the coordinate of the curve, four different cut points (0.5, 1.0, 1.5 and 2.0) were analyzed, resulting in sensitivity rates ranging from 86.2 to 34.5% % and specificity between 87% and 26.1%. By comparing the serum GM index in group case as the presence or absence of antibodies detected by AGID was found p=0.503. The detection of GM the Platelia Aspergillus EIA® test seems is not be useful for the diagnosis of aspergillosis in naturally infected penguins.

Resumo Ainda hoje, um teste diagnóstico eficaz para aspergilose em pinguins não é conhecido, sendo o padrão-ouro os exames post-mortem. O antígeno fúngico galactomanana (GM) tem sido utilizado como biomarcador da doença em humanos, sendo detectado pelo Platelia Aspergillus EIA (BioRad)®, um kit comercial que se baseia na técnica ELISA sanduíche. É padronizado para utilização em pacientes neutropênicos, no entanto estudos tem demonstrado sua possível utilidade também para aves.O objetivo de nosso estudo foi avaliar a eficácia do teste Platelia Aspergillus EIA® (BioRad-US) no diagnóstico da aspergilose em pinguins-de-Magalhães, determinando sensibilidade, especificidade e valores preditivos positivos e negativos em diferentes pontos de corte. Foram incluídas no estudo, amostras de soro sanguíneo (n=29) de pinguins-de-Magalhães em cativeiro que vieram a óbito por aspergilose. A detecção de GM foi realizada seguindo instruções do fabricante e o índice de GM foi obtido dividindo o valor da média da DO da duplicata da amostra clínica pelo valor da média da DO da duplicata da amostra de cut-off fornecida pelo kit. Através de informações em banco de dados foram obtidos resultados sobre a presença de anticorpos anti-Aspergillus fumigatus, detectada por Imunodifusão em gel de ágar (IDGA) em todas as amostras séricas. Os resultados foram analisados utilizando-se teste de qui-quadrado e Kruskal-Wallis a partir do programa estatístico SPSS 20.0, IBM®. Curva ROC foi obtida e a partir desta, taxas de sensibilidade, especificidade, valores preditivo positivo e negativo foram igualmente calculados considerando quatro diferentes pontos de corte (0.5, 1.0, 1.5 e 2.0). O índice de GM sérica não diferiu entre os animais do grupo caso e controle (pKW = 0.097). Na determinação da curva ROC para detecção de GM sérica o valor da área sobre a curva foi de 0.635. A partir dos valores determinados pelas coordenadas da curva, quatro diferentes pontos de corte (0.5, 1.0, 1.5 e 2.0) foram analisados, resultando em taxas de sensibilidade variando de 86.2% a 34.5%, e de especificidade entre 87% e 26.1%. Ao comparar o índice de GM sérica nos animais do grupo caso quanto a presença ou não de anticorpos detectados pela IDGA foi encontrado p=0.503. A detecção de GM pelo teste Platelia Aspergillus EIA® não parece ser útil para o diagnóstico da aspergilose em pinguins naturalmente infectados.

Aislamiento de Aspergillus tritici desde ambiente interno (Chile): alcances ecológicos y clínicos / Isolation of Aspergillus tritici from internal environment (Chile): Ecological and clinical scope

Los ambientes internos son un hábitat protector importante, donde el hombre reside o trabaja la mayor parte de su tiempo. Muchos de estos ambientes carecen de buena ventilación, lo que influye en la composición de sus comunidades microbianas y, en especial, de la fúngica. El objetivo de este estudio es comunicar la presencia de Aspergillus de la sección Candidi en ambientes internos de la Escuela de Medicina de la Universidad de Valparaíso, Chile, y destacar su rol ecológico y su importancia en micología médica. Se efectuó la clasificación morfofisiológica y molecular de dichos aislamientos. Se realizó un muestreo no volumétrico ambiental en agar papa glucosado (PDA) mediante la exposición de 2 placas en 10 diferentes ambientes internos para seleccionar las especies de Aspergillus. Se efectuaron subcultivos en agar Czapek con extracto de levadura (CYA), agar extracto de malta (MEA) y agar creatina sacarosa (CREA) solo para las especies de esporas blancas, para su identificación morfofisiológica y posteriormente molecular. De 20 muestras analizadas, en solo una se aisló un miembro de Aspergillus perteneciente a la sección Candidi. Sobre la base de sus características morfológicas y moleculares, se clasificó a este aislamiento como Aspergillus tritici Mehrotra & Basu. Se describe su ecología y se discute su importancia médica.

Indoor environments provide important protective habitats for humans, who live or work in them most of the time. Many of these environments lack ventilation, which affects the composition of microbial communities, especially that of the fungal community. The aim of this study is to report the isolation of Aspergillus section Candidi from indoor environments of the School of Medicine at Universidad de Valparaiso, Chile, and identification through morpho-physiological and molecular approaches. Their ecological and clinical features were highlighted. An environmental non-volumetric sampling was performed on PDA medium; 2 petri dishes were exposed in 10 different places to select the Aspergillus samples. Subcultures were performed on agar Czapek with yeast extract (CYA), malt extract agar (MEA) and creatin sacarose agar (CREA) media only for the morpho-physiological and later the molecular identification of white spore species. Of the 20 samples analyzed, one Aspergillus belonging to Candidi section was isolated. Based on its morphology and molecular features, it was classified as Aspergillus tritici Mehrotra & Basu. Its ecology and medical relevance are reviewed and discussed.

Assessment of pectinase-producing fungi isolated from soil and the use of orange waste as a substrate for pectinase production

Pectinases are important enzymes not only for their potential applications in different industries such animal feed, agricultural, textile, beverage, food processing, oil extraction, etc. Ten fungal species were isolated from the soil and screened for production of pectinase enzyme by using the pectin agar medium. Pectinolytic enzymes synthesis were attained at a temperature of 30 °C and activities were determined after a seven-days culture of Aspergillus sp. 391 and Aspergillus sp. 031, in a basic medium containing 2% citrus pectin and as the sole carbon source. The extract enzymatic showed an optimum activity for exo-polygalacturonase (PG) and pectin lyase (PNL) against galacturonic acid and pectin at pH 4.5 and 5.5, respectively. There were variations in PG and PNL enzymes levels produced in culture filtrates obtained of Aspergillus sp. 391 with addition of citrus waste (2.0 and 4.0 % w/v) to the medium. Maximum activity for PNL activity was observed in the medium containing 5% pectin or 4% citrus waste, as sole carbon source, after 7 days of growth. The results showed that the isolate Aspergillus sp. 391 is a promising for pectinolytic enzymes production at the industrial level.(AU)

Identification of Fungal Pathogens in Otomycosis and Their Drug Sensitivity: Our Experience

Abstract Introduction Otomycosis is a common problem in otolaryngology practice. However, we usually encounter some difficulties in its treatment because many patients show resistance to antifungal agents, and present high recurrence rate. Objectives To determine the fungal pathogens that cause otomycosis as well as their susceptibility to the commonly used antifungal agents. Additionally, to discover the main reasons for antifungal resistance. Methods We conducted an experimental descriptive study on 122 patients clinically diagnosed with otomycosis from April 2016 to April 2017. Aural discharge specimens were collected for direct microscopic examination and fungal culture. In vitro antifungal susceptibility testing was performed against the commonly used antifungal drugs. We tested the isolated fungi for their enzymatic activity. Results Positive fungal infection was found in 102 samples. The most common fungal pathogens were Aspergillus and Candida species, with Aspergillus niger being the predominant isolate (51%). The antifungal susceptibility testing showed that mold isolates had the highest sensitivity to voriconazole (93.48%), while the highest resistance was to fluconazole (100%). For yeast, the highest sensitivity was to nystatin (88.24%), followed by amphotericin B (82.35%), and the highest resistance was to terbinafine (100%), followed by Itraconazole (94.12%). Filamentous fungi expressed a high enzymatic ability, making them more virulent. Conclusion The Aspergillus and Candida species are the most common fungal isolates in otomycosis. Voriconazole and Nystatin are the medications of choice for the treatment of otomycosis in our community. The high virulence of fungal pathogens is owed to their high enzymatic activity. Empirical use of antifungals should be discouraged.

The occurrence of aflatoxigenic Aspergillus spp. in dairy cattle feed in Southern Brazil

ABSTRACT The presence of mycotoxins or related fungi in animal feed is a major problem for animal and human health. Silage and concentrated feed samples were collected from 21 dairy farms in the Western part of Paraná state in Southern Brazil. Water activity and pH of all samples were measured, and each sample was analyzed to check for the presence of aflatoxigenic Aspergillus. Water activity was observed to be lower in the concentrated feed samples. The pH was lower in the silage samples, indicating fermentation processes. Two silage samples and four concentrated feed samples were contaminated with Aspergillus spp. Seven isolates of Aspergillus spp. were obtained and their potential to produce aflatoxins was evaluated. Four of the isolates, two from the silage samples and two from the concentrated feed samples, produced the aflatoxins B1, B2, G1, and G2 in culture media. These isolates were identified as Aspergillus parasiticus and Aspergillus nomius. The presence of aflatoxigenic isolates of Aspergillus spp. in silage and concentrated feed samples is a matter of concern, because of the risk of aflatoxin production and contamination of the animal feed.

Comparison of the recovery rate of otomycosis using betadine and clotrimazole topical treatment / Comparação da taxa de recuperação de otomicose com o uso de tratamento tópico com betadina e clotrimazol

Abstract Introduction Otomycosis is a common diseases that can be associated with many complications including involvement of the inner ear and mortality in rare cases. Management of otomycosis can be challenging, and requires a close follow-up. Treatment options for otomycosis include local debridement, local and systemic antifungal agents and utilization of topical antiseptics. Objective This study was designed to compare the recovery rate of otomycosis using two therapeutic methods; topical betadine (Povidone-iodine) and clotrimazole. Methods In this single-blind clinical trial, 204 patients with otomycosis were selected using a non-probability convenient sampling method and were randomly assigned to two treatment groups of topical betadine and clotrimazole (102 patients in each group). Response to treatment was assessed at 4, 10 and 20 days after treatment. Data were analyzed using the independent t-test, Chi-Square and Fisher exact test in SPSS v.18 software, at a significance level of p < 0.05. Results The results showed that out of 204 patients with otomycosis, fungi type isolated included Aspergillus in 151 cases (74%), and Candida albicans in 53 patients (26%). On the fourth day after treatment, 13 patients (13.1%) in the group treated with betadine and 10 patients (9.8%) in the group treated with clotrimazole showed a good clinical response to treatment (p = 0.75). A good response to treatment was reported for 44 (43.1%) and 47 patients (46.1%) on the tenth day after the treatment (p = 0.85); and 70 (68.6%) and 68 patients (67.6%) on the twentieth day after treatment (p = 0.46) in the groups treated with betadine and clotrimazole, respectively. The response to treatment was thus not significantly different in the two groups. Conclusion In the present study the efficacy of betadine and clotrimazole was the same for the treatment of otomycosis. The result of this study supports the use of betadine as an effective antifungal in otomycosis treatment, helping to avoid the emergence of resistant organisms.

Resumo Introdução A otomicose é uma das doenças comuns associadas a muitas complicações, como envolvimento da orelha interna e mortalidade em casos raros. O tratamento da otomicose pode ser realmente desafiador e requer um acompanhamento rigoroso. As opções de tratamento para otomicose podem incluir desbridamento local, agentes antifúngicos locais e sistêmicos e uso de antissépticos tópicos, os medicamentos tópicos recomendados para o tratamento da otomicose. Objetivo Comparar a taxa de recuperação de otomicose utilizando dois métodos terapêuticos de betadina tópica (povidona-iodo) e clotrimazol. Método Neste ensaio clínico simples cego, 204 pacientes com otomicose foram selecionados utilizando-se método de amostragem de não probabilidade conveniente e randomizados para dois grupos de tratamento, com betadina tópica e com clotrimazol (102 pacientes em cada grupo). A resposta ao tratamento foi avaliada aos 4, 10 e 20 dias após o tratamento. Os dados foram analisados utilizando o teste t independente, qui-quadrado e teste de Fisher no software SPSS v.18, com nível de significância de p < 0,05. Resultados Os resultados mostraram que dos 204 pacientes com otomicose, os tipos de fungos isolados incluíram Aspergillus em 151 casos (74%) e Candida albicans em 53 pacientes (26%). No quarto dia após o tratamento, 13 pacientes (13,1%) no grupo tratado com betadina e 10 pacientes (9,8%) no grupo tratado com clotrimazol apresentaram boa resposta ao tratamento (p = 0,75). Uma boa resposta ao tratamento foi relatada para 44 (43,1%) e 47 pacientes (46,1%) no décimo dia após o tratamento (p = 0,85); e 70 (68,6%) e 68 pacientes (67,6%) no vigésimo dia após o tratamento (p = 0,46) no grupo tratado com betadina e clotrimazol, respectivamente. Assim, a resposta ao tratamento não foi significativamente diferente nos dois grupos. Conclusão No presente estudo, a eficácia da betadina e do clotrimazol foi a mesma no tratamento da otomicose. O resultado deste estudo apoia o uso de betadina como um antifúngico eficaz no tratamento da otomicose que pode ajudar a evitar o surgimento de organismos resistentes.

Biodiversity of species of Aspergillus section Fumigati in semi-desert soils in Argentina / Biodiversidad de especies de Aspergillus de la sección Fumigati en suelos semidesérticos de Argentina

The distribution of Aspergillus species in soil has been widely studied all over the world. The aim of this study was the phenotypic and genotypic characterization of species belonging to section Fumigati present in soils from two Argentinian semi-desert areas having different geological conditions. Altogether, 23 isolates belonging to Aspergillus section Fumigati were recovered and identified using a polyphasic approach including phenotypic and molecular identifications. Aspergillus fumigatus sensu stricto and Aspergillus fumigatiaffinis had the highest frequency, of occurrence while isolates closely related to Aspergillus udagawae and Aspergillus felis were rarely observed. A. fumigatiaffinis and isolates closer to A. udagawae were isolated for the first time from Argentinian soils and this is the first report on the occurrence of species belonging to the A. felis clade in South America. Recent scientific interests in biodiversity, as well as the increasing importance of aspergilli as causative agents of human and animal diseases increase the need to understand the diversity and occurrence of these fungi in nature.

La distribución de especies de Aspergillus en el suelo se ha estudiado ampliamente en todo el mundo. El objetivo de este trabajo fue caracterizar fenotípica y genotípicamente las especies pertenecientes a la sección Fumigati presentes en los suelos de dos zonas semidesérticas de Argentina con diferentes geologías. En total, 23 Aspergillus de la sección Fumigati fueron aislados e identificados utilizando un enfoque polifásico incluyendo identificaciones fenotípicas y moleculares. Aspergillus fumigatus sensu stricto y Aspergillus fumigatiaffinis aparecieron con mayor frecuencia, mientras que los aislamientos relacionados a Aspergillus udagawae y a Aspergillus felis se observaron raramente. Este es el primer informe de A. fumigatiaffinis y de aislamientos estrechamente relacionados a A. udagawae en suelos argentinos; también el primero sobre la ocurrencia de especies pertenecientes al clado A. felis en Sudamérica. El emergente interés científico en la biodiversidad, así como la creciente importancia de Aspergillus como agentes causales de enfermedades humanas y animales, aumentan la necesidad de conocer la diversidad y la ocurrencia de estos hongos en la naturaleza.

Fungos potencialmente toxigênicos em amostras de amendoim disponível para o consumo humano / Potentially toxigenic fungi in peanut samples available for human consumption

Avaliou-se a contaminação por fungos potencialmente toxigênicos em amendoim in natura disponível para consumo humano e comercializado em supermercados de Juiz de Fora, MG. Foram adquiridas 31 amostras de sete diferentes marcas de amendoim em grãos cru; amendoim torrado em grãos e amendoim torrado e triturado (moído), em embalagens originais e invioladas, em cinco estabelecimentos comerciais. A análise da presença de fungos potencialmente toxigênicos nos grãos de amendoim foi realizada por meio da técnica de plaqueamento direto em placas de Petri, contendo os meios de cultura Ágar Batata Dextrose (ABD) e Ágar Dicloran Rosa de Bengala Cloranfenicol (ADRBC). Das 31 amostras analisadas, 17 (54,84%) estavam contaminadas por fungos potencialmente toxigênicos (Aspergillus niger, Aspergillus flavus e/ou Aspergillus parasiticus, Aspergillus fumigatus e Penicillium spp) além de outros não toxigênicos (Rhizopus spp. e leveduras). O índice de contaminação nas diferentes amostras de amendoim avaliadas foi expressivo, sendo que as espécies encontradas foram de fungos potencialmente toxigênicos produtores de micotoxinas importantes como as aflatoxinas.

Invasive fungal infection in patients with hematologic disorders in a Brazilian tertiary care hospital

ABSTRACT INTRODUCTION: Invasive fungal infections (IFIs) are an important complication in immunocompromised individuals, particularly neutropenic patients with hematological malignancies. In this study, we aimed to verify the epidemiology and diagnosis of IFIs in patients with hematologic problems at a tertiary hospital in Goiânia-GO, Brazil. METHODS: Data from 117 patients, involving 19 cases of IFIs, were collected. The collected data included diagnosis methods, demographics, clinical characteristics, and in vitro susceptibility to different antifungal agents. Among the 19 cases, 12 were classified as proven IFI and 7 as probable invasive aspergillosis with detection of galactomannan in blood and presence of lung infiltrates in radiographic images. Logistic regression analysis showed that the proven and probable IFIs were associated with increased risk of death. Statistical analysis demonstrated that age, sex, and underlying disease were not independently associated with risk of death in IFI patients. RESULTS: Most bloodstream isolates of Candida spp. exhibited low minimum inhibitory concentrations (MICs) to all antifungal agents tested. Voriconazole and amphotericin had the lowest MICs for Aspergillus spp. and Fusarium spp., but Fusarium spp. showed the least susceptibility to all antifungals tested. Amphotericin B, fluconazole, and itraconazole were found to be inactive in vitro against Acremonium kiliense; but this fungus was sensitive to voriconazole. CONCLUSIONS: Considering the high number of IFI cases, with crude mortality rate of 6%, we could conclude that IFIs remain a common infection in patients with hematological malignancies and underdiagnosed ante mortem. Thus, IFIs should be monitored closely.

Síntese de heteroaril oxazolinas e derivados triazólicos com potencial atividade biológica / Synthesis of heteroaryl oxazolines and triazolic derivatives with potential biological activity

Fungos no ambiente podem ser patogênicos ou oportunistas, dependendo da imunidade do hospedeiro. Existem várias espécies de fungos, por exemplo, Cândida albicans, Cryptococcus e Aspergillus. A primeira espécie fúngica pode ser tratada com o antifúngico fluconazol, que é um composto que contém anéis heterocíclicos 1,2,4-triazólicos. Além disso, existem cepas de fungos que são resistentes à terapia com fluconazol, que é o caso das Cândida krusei, Cândida tropicalis; entre outras. A busca por novos tratamentos envolve o desenvolvimento de novas moléculas sintéticas. Neste trabalho, sintetizamos uma biblioteca de compostos oxazolínicos e seus derivados 1,2,3-triazólicos. A atividade microbiológica foi avaliada contra 10 tipos de Cândida, 2 tipos de Cryptococcus e 2 tipos de Aspergillus. Além disso, foram feitos os testes de hemólise, citotoxicidade, combinações de drogas e permeabilidade de membrana. Os resultados sugerem um alto potencial terapêutico dos compostos e os propomos como potenciais novos antifúngicos

ungi in the environment may be pathogenic or opportunistic depending on the immune status of the host. There are several species of fungi, for example, Candida albicans, Cryptococcus and Aspergillus. The first fungal species can be treated with the antifungal fluconazole, which is a compound containing 1,2,4-triazole heterocyclic rings. In addition, there are strains of fungi that are resistant to fluconazole therapy, which is the case of Candida krusei, Candida tropicalis; among others. The search for new treatments involves the development of new synthetic molecules. In this work, we synthesized a library of oxazoline compounds and their 1,2,3-triazole derivatives. Microbiological activity was evaluated against 10 types of Candida, 2 types of Cryptococcus and 2 types of Aspergillus. In addition, hemolysis, cytotoxicity, drug combinations and membrane permeability were performed. The results suggest the high therapeutic potential of the compounds and we propose them as potential new antifungals

Aspergilosis invasora post-quirúrgica: presentación de caso clínico / Invasive aspergillosis post surgical: presentation of case

La aspergilosis invasora es una infección oportunista causada por hongos del género Aspergillus spp, considerándose la con peor pronóstico producida por este organismo. Presenta una elevada tasa de mortalidad a pesar del avance en los tratamientos antifúngicos, afectando principalmente a pacientes inmunosuprimidos. Dentro de las formas clínicas se encuentra la que sucede posterior a una cirugía conllevando una elevada morbi-mortalidad. Presentamos un caso clínico del Hospital Roberto Del Río. Recién nacido de término diagnosticado de transposición de grandes vasos, quien a los 9 días de vida es sometido a cirugía correctora. Egresa de pabellón con tórax abierto y con requerimiento de drogas vasoactivas. Al décimo día postoperatorio presenta deterioro hemodinámico, se realiza ecocardiograma que muestra líquido con ecorefringencias y se realiza aseo quirúrgico, extrayendo muestra para cultivo que resulta positivo para Aspergillus terreus. Por consiguiente, se inicia tratamiento con voriconazol con buena respuesta clínica...

Invasive aspergillosis is an opportunistic infection caused by fungi of the genus Aspergillus spp, considered the worst prognosis produced by this organism. It has a high mortality despite progress in antifungal treatments, affecting mainly immunocompromised patients. Clinically can occur following surgery , leading to high morbidity and mortality. We report a case of Roberto Del Río Hospital. Term newborn diagnosed with transposition of the great vessels, who at 9 days of life undergoes corrective surgery. Leaves the pavilion with open chest and requirement of vasoactive drugs. On the tenth day after surgery presents hemodynamic deterioration, the echocardiogram shows refringence and surgical cleaning is performed, a sample is removed and cultured resulting positive for Aspergillus terreus. Therefore, treatment with voriconazole starts with good clinical response...

Infecciones fúngicas en pacientes hemato-oncológicos pediátricos / Fungal infections in pediatric hematology-oncology patients

Introducción: Las infecciones fúngicas invasivas (IFI) son una causa importante de morbimortalidad en pacientes inmunocomprometidos. En las últimas décadas se produjo un incremento, variaciones epidemiológicas y avances en los métodos de diagnóstico y tratamiento de las mismas. El objetivo de este estudio fue analizar las características epidemiológicas, clínicas y de evolución de IFI en pacientes hematooncológicos. Material y Métodos: Estudio de cohorte observacional retrospectivo y prospectivo. Se incluyeron pacientes con edades entre 1 mes y 18 años con diagnóstico de enfermedad hemato-oncológica admitidos en Hospital Juan P. Garrahan en el período 01/2010 - 04/2014 con diagnóstico de IFI según la EORTC. Resultados: Durante el período de estudio se incluyeron 124 pacientes con IFI. La incidencia fue de 2,65 casos /100 episodios febriles. Las enfermedades de base correspondieron a leucemias agudas en 66,1% (n:82) y a trasplante de médula ósea en 27,4%. Los períodos de mayor riesgo de aparición de IFI fueron las etapas de inducción (21,8%), recaída (16,9%) y re- inducción (12,9%). En general las IFI (n: 110; 88,7%) ocurrieron en el contexto de neutropenia febril. La documentación microbiológica demostró el predominio de Aspergillus spp. especies de Candida no albicans y baja prevalencia de mucorales. Se evidenció co-infección en 80 pacientes (64,5%). El tratamiento antifúngico empírico fue anfotericina liposomal en 48,8% de los pacientes, 46,3% recibió anfotericina desoxicolato y 4,9% voriconazol. Ingresaron a la UCI 30 pacientes (31,5%). La evolución de los pacientes con IFI fue favorable en 77,4%, de los casos; mientras que fallecieron 28 (22,6%). Del total de los pacientes fallecidos, 23 (82%) tuvieron una infección concomitante. Conclusiones: La incidencia de IFI documentada en nuestro estudio fue de 2,65 casos /100 episodios febriles, las leucemias agudas fueron las patologías de base más frecuentemente asociadas y la mayoría de las IFI se asoció a neutropenia febril. Aspergillus spp. fue el hongo más frecuentemente hallado. La mortalidad relacionada a IFI fue de 22,6% La presencia de co-infecciones se asoció con peor evolución (AU)

Introduction: Fungal invasive infections (FII) are an important cause of morbidity and mortality in immunocompromised patients. Over the past decades an increase in incidence, epidemiologic variations, diagnostic methods, and treatment has occurred. The aim of this study was to analyze epidemiological and clinical features and outcome of FII in hematology-oncology patients. Material and Methods: An observational retrospective and prospective cohort study. Patients aged between 1 month and 18 years with hematology-oncology disease admitted to Hospital Juan P. Garrahan from 01/2010 to 04/2014 diagnosed with FII according to the EORTC were included. Results: During the study period 124 patients with FII were included. Incidence was 2,65 cases /100 febrile episodes. Underlying diseases were acute leukemia in 66,1% (n:82) and bone marrow transplantation in 27,4%. The periods of increased risk of FII were the induction (21,8%), relapse (16,9%), and re-induction (12,9%) stages. FII occurred typically in the context of febrile neutropenia (n: 110; 88,7%). Microbiology predominantly showed Aspergillus spp, non-albicans Candida spp, and a lower prevalence of mucor sp. Co-infection was observed in 80 patients (64,5%). Empirical antifungal therapy was liposomal amphotericin in 48.8% of the patients, amphotericin B deoxycholate in 46.3%, and voriconazole in 4,9%. Thirty patients (31,5%) were admitted to the ICU. Outcome of patients with FII was favorable in 77,4% of the cases, while 28 (22,6%) died. Of all patients that died 23 (82%) had a concomitant infection. Conclusions: In our study, incidence of documented FII was 2,65 cases /100 febrile episodes. Acute leukemia was the most common underlying disease and the majority of FII were associated with febrile neutropenia. Aspergillus spp was the most commonly found fungus. FII-related mortality was 22,6%. The presence of coinfections was associated with worse outcome (AU)

Factores extrínsecos e intrínsecos asociados a poblaciones fúngicas micotoxigénicas de granos de maíz (Zea mays L) almacenados en silos bolsa en Argentina / Extrinsic and intrinsic factors associated with mycotoxigenic fungi populations of maize grains (Zea mays L.) stored in silobags in Argentina

Con el objeto de caracterizar las poblaciones fúngicas, en particular las especies potencialmente micotoxigénicas, que pueden contaminar los granos de maíz almacenados en silos bolsa con un contenido de humedad superior al recomendado como seguro, se evaluaron 270 muestras extraídas al inicio, a los 90 días y al final de un período de almacenamiento de 5 meses. En dichas muestras se cuantificó e identificó la biota fúngica y se determinó la contaminación con fumonisinas y aflatoxinas. Asimismo, se evaluó el efecto de factores extrínsecos (ambiente), intrínsecos (granos) y tecnológicos (ubicación de los granos en el perfil del silo bolsa) sobre las poblaciones totales y micotoxigénicas. El pH de los granos y el nivel de O2 se redujeron significativamente a los 5 meses, mientras que la concentración de CO2 se incrementó en igual período. Los recuentos totales de la micobiota fueron significativamente mayores en los granos ubicados en el estrato superior del silo bolsa. Se identificaron especies micotoxigénicas de Fusarium, Aspergillus, Penicillium y Eurotium. La frecuencia de aislamiento de Fusarium verticillioides se redujo al final del almacenamiento y Aspergillus flavus solo se aisló en el inicio del almacenamiento. Los recuentos de Penicillium spp. y Eurotium spp. se incrementaron al final del almacenamiento. El 100 % de las muestras presentaron contaminación con fumonisinas, con niveles máximos de 5,707 mg/kg, mientras que las aflatoxinas contaminaron el 40 % de las muestras con niveles máximos de 0,0008 mg/kg. Las condiciones ambientales y de sustrato generadas durante el almacenamiento produjeron cambios en la composición de las poblaciones fúngicas y limitaron el desarrollo de hongos micotoxigénicos y la producción de micotoxinas.

In order to determine the behavior of mycotoxin-producing fungal populations linked with silobags stored corn grains with a moisture content greater at the recommended as safe, 270 samples taken in three times (beginning, 90 days, final) over a five month period of storage were evaluated. The fungal biota was quantified and identified and the contamination with fumonisin and aflatoxin was determined. Extrinsic factors (environment), intrinsic factors (grains) and technological factors (location of the grains in the profile of silobag) were taken into account to evaluate the presence and quantity of total and mycotoxigenic fungal populations. The pH of grains and O2 levels were significantly reduced after five months, while CO2 concentration increased in the same period. The total counts of mycobiota were significantly higher in grains located in the top layer of silobag. Mycotoxigenic species of Fusarium, Aspergillus, Penicillium and Eurotium were identified. The frequency of isolation of Fusarium verticillioides decreased at the end of storage and Aspergillus flavus was isolated only at the beginning of storage. The counts of the Penicillium spp. and Eurotium spp. were increased at the end of storage. Fumonisin contamination was found in all the samples (100 %) with maximum levels of 5.707 mg/kg whereas aflatoxin contaminated only 40 % with maximum levels of 0.0008 mg/kg. The environmental and substrate conditions generated during the storage limited the development of mycotoxigenic fungi and mycotoxin production.

Utilidad de la prueba Aspergillus-LFD para el diagnóstico de aspergilosis: primera experiencia en Chile / Utility of Aspergillus-LFD: first experience in Chile

Introduction: Invasive fungal diseases (IFD) by filamentous fungi are a common cause of morbidity and mortality in immunocompromised patients, especially those with myeloid leukemia. In 2011 a protocol for the rapid diagnosis of IFD by filamentous fungi was implemented in Valparaiso Region. Objectives: To describe cases of IFD by filamentous fungi of the Valparaíso Region, since the implementation of rapid diagnosis and to compare results with the period 2004-2009. Materials and Method: Descriptive and prospective study conducted in two public hospitals: Carlos van Buren at Valparaiso and Gustavo Fricke at Viña del Mar. We selected patients with a diagnosis of filamentous fungal diseases considering the EORTC/MSG criteria. Demographics, underlying diseases, risk factors for EFI, galactomannan (GM) results in blood and bronchoalveolar lavage, cultures and biopsies, treatment and overall lethality rates at 30 days were registered. Results: Eighteen patients were detected, 6 with proven and 12 probable IFD. Nine were diagnosed by GM, 8 by culture and two with both methods. In cases which the agent (9/18) was isolated from Rhizopus oryzae was the most frequent. When comparing overall lethality with the period 2004-2009, there was a reduction of 47.8%, which was statistically significant. Conclusions: Compared to data previously published in the region, demographic and comorbidities of patients with IFD caused by filamentous fungi are similar, however the currently rapid diagnosis protocol has improved survival of patients and lethality experienced overall decrease.

Introducción: la enfermedad fúngica invasora (EFI) por hongos filamentosos es una causa frecuente de morbilidad y mortalidad en pacientes inmunocomprometidos, en especial en aquellos con leucemia mieloide. En el 2011 se implementó en la Región de Valparaíso un protocolo de diagnóstico rápido de la EFI por hongos filamentosos. Objetivos: describir los casos de EFI por hongos filamentosos de la Región de Valparaíso, desde la implementación del diagnóstico rápido y compararlos con el período 2004-2009. Materiales y Método: Estudio descriptivo y prospectivo realizado en los hospitales públicos Carlos van Buren de Valparaíso y Gustavo Fricke de Viña del Mar. Se seleccionaron aquellos pacientes con diagnóstico de EFI por hongos filamentosos considerando los criterios EORTC/MSG. Se obtuvieron datos demográficos, enfermedad de base, factores de riesgo para EFI, resultados de galactomanano (GM), cultivos y biopsias, tratamiento y letalidad global a 30 días. Resultados: Se identificaron 18 pacientes, seis con EFI probadas y 12 probables. Nueve fueron diagnosticados con galactomanano, ocho con cultivos y uno con los dos métodos. En los casos en que se aisló el agente (9/18), Rhizopus oryzae fue el más frecuente. Al comparar la letalidad global con la del período 2004-2009, hubo una reducción de 47,8%, la cual fue estadísticamente significativa. Conclusiones: En relación a lo publicado anteriormente en la región, se conservan las características demográficas y de co-morbilidad de los pacientes con EFI por hongos filamentosos; sin embargo, la introducción del nuevo protocolo de diagnóstico rápido se asoció a una disminución en la letalidad global.

Invasive Primary Colonic Aspergillosis in the Immunocompetent Host without Classical Risk Factors

Invasive aspergillosis (IA), generally considered an opportunistic infection in immunocompromised hosts, is associated with high morbidity and mortality. IA commonly occurs in the respiratory tract with isolated reports of aspergillosis infection in the nasal sinuses, central nervous system, skin, liver, and urinary tract. Extra-pulmonary aspergillosis is usually observed in disseminated disease. To date, there are a few studies regarding primary and disseminated gastrointestinal (GI) aspergillosis in immunocompromised hosts. Only a few cases of primary GI aspergillosis in non-immunocompromised hosts have been reported; of these, almost all of them involved the upper GI tract. We describe a very rare case of IA involving the lower GI tract in the patient without classical risk factors that presented as multiple colon perforations and was successfully treated by surgery and antifungal treatment. We also review related literature and discuss the characteristics and risk factors of IA in the immunocompetent hosts without classical risk factors. This case that shows IA should be considered in critically ill patients, and that primary lower GI aspergillosis may also occur in the immunocompetent hosts without classical risk factors.

Recent Clinical Manifestation and Prognosis of Fungal Endophthalmitis: A 7-Year Experience at a Tertiary Referral Center in Korea

This study analyzed the recent causes, prognosis, and treatment strategies for fungal endophthalmitis. A retrospective review of patients who were diagnosed with fungal endophthalmitis at our center was conducted. The fungal organisms isolated from each patient and the visual prognosis according to the route of infection and treatment method were analyzed. A total of 40 eyes from 30 patients with fungal endophthalmitis were included in this study. Candida species were the most common causative organisms in 35 of 40 eyes. Endogenous and exogenous endophthalmitis were observed in 33 and 7 eyes, respectively. Pre- and post-treatment best-corrected visual acuity (BCVA) was not significantly different between endogenous endophthalmitis and exogenous endophthalmitis. The 40 eyes were treated using the following modalities: intravitreal antifungal agent injection with intravenous antifungal agent (16 eyes), vitrectomy with intravenous antifungal agent (14 eyes), intravenous antifungal agent alone (9 eyes), and evisceration (1 eye). Post-treatment BCVA only significantly improved after treatment in the vitrectomy group. Candida species were the most common cause of fungal endophthalmitis, irrespective of the route of infection. The visual prognosis of fungal endophthalmitis was generally poor. In conclusion, if the general condition of the patient tolerates a surgical procedure, prompt vitrectomy and intravitreal injection of antifungal agents can improve visual acuity.

Fungi in the respiratory tract of horses with recurrent airway obstruction / Presença de fungos no trato respiratório de equinos e sua associação com obstrução recorrente das vias aéreas

This study aims to compare the concentration of viable fungi, especially those of the genus Aspergillus in the respiratory tract of stabled horses with and without Recurrent Airway Obstruction (RAO). Thirty two housed horses from four equestrian centers in Brazil were included in the study. These animals were submitted to clinical examination and to a respiratory sample collection. They were categorized into two groups: healthy and RAO-affected horses. Samples obtained by tracheobronchial washes were evaluated for fungal microscopy, quantitative culture and Aspergillus spp. quantification. Eighteen healthy and 14 RAO-affected horses were studied. Fungi were more frequently recovered in the RAO group, in comparison to controls, for both fungal microscopy (P<0.0001), fungal culture (P<0.0001) and Aspergillus spp. quantitative culture (p=0.001). In conclusion, horses with RAO have significantly higher fungal load in the respiratory tract in comparison to healthy horses. The implications of these findings in terms of the pathogenesis of RAO deserve additional investigation...

Este estudo objetivou comparar a concentração de fungos viáveis, especialmente do gênero Aspergillus, no trato respiratório de equinos estabulados com e sem obstrução recorrente das vias aéreas (ORVA). Trinta e dois equinos provenientes de quatro centros de treinamento equestre do Brasil foram incluídos no estudo. Os animais foram submetidos a exame clínico e coleta de amostra respiratória, sendo categorizados em dois grupos: sadios e ORVA. Os lavados traqueobrônquicos obtidos foram avaliados por exame micológico direto e cultivo quantitativo. Ao todo, 18 equinos saudáveis e 14 com ORVA foram estudados. Fungos foram mais frequentemente detectados em amostras do grupo ORVA em comparação com o grupo controle, tanto no exame micológico direto (P<0,0001) e cultivo quantitativo (P<0,0001) quanto na concentração de unidades formadoras de colônias (UFC) de Aspergillus spp. isolada em cultivo (p=0,001). Equinos com ORVA têm maior concentração de propágulos fúngicos no trato respiratório em comparação com animais sadios. As implicações desses achados na patogenia da ORVA merecem maior atenção e investigação...