Molecular detection of Paracoccidioides spp. in environmental samples from the Southwestern Amazon.

Paracoccidioidomycosis is an infection with the potential for environmental dissemination, especially in regions of hot and humid climate, where human cases have been recorded in the Southwestern Amazon of Brazil, specifically in the state of Acre. Despite studies providing information about the presence of these fungi in soil and animal samples, such as armadillos, further investigations are still needed to determine the epidemiological distribution of the genus Paracoccidioides. The aim of this study was to detect the occurrence of Paracoccidioides fungi in the Southwestern Amazon. To achieve this, 60 soil samples were collected from armadillo burrows on rural properties in the in the municipalities of Acrelândia, Bujari, Plácido de Castro, Rio Branco, Sena Madureira, and Senador Guiomard, located in the state of Acre, Brazil. Fungal DNA was extracted from these samples using the DNEASY® PowerSoil kit-Quiagen, followed by Nested PCR technique with ITS4 and ITS5 as external primers, and PBITS-E and PBITS-R as internal primers. DNA amplification products of about 380 bp compatible with Paracoccidioides spp. were detected in six samples (10%), being sequenced and identified as P. brasiliensis. These findings indicate that the soils of the Acre state could be considered a potential source for Paracoccidioides spp., suggesting that local infections are likely.

Oral glutamine supplementation relieves muscle loss in immobilized rats, altering p38MAPK and FOXO3a signaling pathways.

BACKGROUND: Skeletal muscle synthesizes, stores, and releases body L-glutamine (GLN). Muscle atrophy due to disabling diseases triggers the activation of proteolytic and pro-apoptotic cell signaling, thus impairing the body's capacity to manage GLN content. This situation has a poor therapeutic prognosis. OBJECTIVE: Evaluating if oral GLN supplementation can attenuate muscle wasting mediated by elevated plasma cortisol and activation of caspase-3, p38MAPK, and FOXO3a signaling pathways in soleus and gastrocnemius muscles of rats submitted to 14-day bilateral hindlimbs immobilization. METHODS: Animals were randomly distributed into six groups: non-immobilized rats (Control), control orally supplemented with GLN (1 g kg-1) in solution with L-alanine (ALA: 0.61 g kg-1; GLN+ALA), control orally supplemented with dipeptide L-alanyl-L-glutamine (DIP; 1.49 g kg-1), hindlimbs immobilized rats (IMOB), IMOB orally GLN+ALA supplemented (GLN+ALA-IMOB), and IMOB orally DIP supplemented (DIP-IMOB). Plasma and muscle GLN concentration, plasma cortisol level, muscle caspase-3 activity, muscle p38MAPK and FOXO3a protein content (total and phosphorylated forms), and muscle cross-sectional area (CSA) were measured. RESULTS: Compared to controls, IMOB rats presented: a) increased plasma cortisol levels; b) decreased plasma and muscle GLN concentration; c) increased muscle caspase-3 activity; d) increased total and phosphorylated p38MAPK protein content; e) increased FOXO3a and decreased phosphorylated FOXO3a protein content; f) reduced muscle weight and CSA befitting to atrophy. Oral supplementation with GLN+ALA and DIP was able to significantly attenuate these effects. CONCLUSIONS: These findings attest that oral GLN supplementation in GLN+ALA solution or DIP forms attenuates rats' skeletal muscle mass wasting caused by disuse-mediated muscle atrophy.

Occurrence, Biological and Enzymatic Activities of Agaricomycetes from the Brazilian Amazon - A Systematic Review.

Agaricomycetes are highlighted for producing a variety of compounds and enzymes with nutritional and medicinal properties. However, the knowledge of the diversity of this group of fungi is still insufficient, as well as their biological and enzymatic activities. Thus, the objective of this work is to describe the occurrence, biological and enzymatic activities of Agaricomycetes from the Brazilian Amazon. The present study is a systematic review with the literature search done in the following databases: Scielo, Google Scholar, PubMed and ScienceDirect. The descriptors used were Basidiomycota, Agaricomycetes, mushroom, antimicrobial activity, antitumor activity, antioxidant activity, anti-inflammatory activity, immunomodulator, enzymatic activity, and Brazilian Amazon. We used as inclusion criteria articles in Portuguese and English, published between 2010 and 2021 and that had the full text available and presented relevance to the exposed topic, and as exclusion criteria, works not done in the Brazilian Amazon, duplicate articles in the databases search or outside the topic under study. A total of 40 articles, published between 2010 and 2021, were selected for analysis. 230 species of Agaricomycetes fungi were described for the Brazilian Amazon, with the most frequent orders being Polyporales (52.60%), Agaricales (14.35%), and Hymenochaetales (13.91%). Six studies were found on antimicrobial activity for promising Agaricomycete fungi against the bacteria Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and the fungi Candida albicans, C. parapsilosis and C. tropicalis. For the antioxidant activity, a study described the species Lentinus citrinus with high amounts of antioxidant compounds. For enzymatic activity, five studies reported Agaricomycete fungi producing protease, cellulase, amylase, pectinase, laccase, and xylanase enzymes. This review shows the scarcity of studies on the description and technological potential of Agaricomycetes from the Brazilian Amazon, highlighting the need to encourage the study of this group of organisms.

Conhecimento dos acadêmicos dos cursos da saúde acerca de vacinação / Knowledge of health course academics about vaccination / Conocimiento de académicos en cursos de salud sobre vacunación

Objetivo: Analisar os fatores associados ao conhecimento de acadêmicos de enfermagem e medicina acerca da vacinação. Método: Trata-se de um estudo descritivo e analítico, transversal, realizado com os acadêmicos dos cursos de Enfermagem e Medicina de uma universidade pública. Os dados foram obtidos através de questionários com perguntas sobre as características demográficas, acadêmicas e relacionadas ao tema vacinação. Os dados foram trabalhados no programa estatístico Stata 16. Resultados: Participaram desta pesquisa 113 estudantes, sendo maioria dos entrevistados do sexo feminino, vinculados ao curso da Enfermagem, com predomínio de idade na faixa etária entre 18 a 23 anos, e que se encontrava no terceiro ano de curso. Foi apontado baixo conhecimento dos acadêmicos, destacando-se uma associação significativa com as variáveis faixa etária, curso acadêmico e anos cursados. Conclusão: É importante salientar o papel da universidade na formação dos profissionais de saúde, pois os mesmos carecem de conhecimento ao longo da formação acadêmica acerca do tema vacinação. (AU).

Objective: To analyze the factors associated with the knowledge of nursing and medical students about vaccination. Method: This is a descriptive and analytical, cross-sectional study carried out with nursing and medical students at a public university. Data were obtained through questionnaires with questions about demographic, academic and vaccination-related characteristics. Data were processed using the Stata 16 statistical program. Results: 113 students participated in this research, the majority of respondents being female, nursing students, with a predominance of age in the age group between 18 and 23 years old, and who were in the third year of course. Low knowledge of academics was pointed out, highlighting a significant association with the variables age group, academic course and years studied. Conclusion: It is important to emphasize the role of the university in the training of health professionals, as they lack knowledge throughout their academic training on the topic of vaccination. (AU).

Objetivo: Analizar los factores asociados al conocimiento de estudiantes de enfermería y medicina sobre vacunación. Método: Se trata de un estudio descriptivo y analítico, de corte transversal, realizado con estudiantes de enfermería y medicina de una universidad pública. Los datos se obtuvieron a través de cuestionarios con preguntas sobre características demográficas, académicas y vacunales. Los datos fueron procesados mediante el programa estadístico Stata 16. Resultados: Participaron de esta investigación 113 estudiantes, siendo la mayoría de los encuestados del sexo femenino, vinculados a la carrera de Enfermería, con predominio de la edad en el grupo etario entre 18 y 23 años, y que estaban en el tercer año, por supuesto. Se señaló bajo conocimiento académico, destacándose asociación significativa con las variables grupo etario, curso académico y años de estudio. Conclusión: Es importante resaltar el papel de la universidad en la formación de los profesionales de la salud, ya que estos carecen de conocimientos a lo largo de su formación académica sobre el tema de la vacunación. (AU).

NATIVE AMAZONIAN FUNGI TO CONTROL TERMITES Nasutitermes sp. (BLATTODEA: TERMITIDAE) / Hongos natives para el control de Nasutitermes sp. (Blattodea: Termitidae)

ABSTRACT Termites of the Nasutitermes genus are considered one of the main urban and agroforestry pests in Brazil, where the main method of control is the application of pesticides. The uso of entomopathogenic fungi to reduce the population of this plague in the environment could be use as an alternative. The goal of this study was to evaluate the virulence of isolates native Amazonian fungi belonging to the Tolypocladium endophyticum, Metarhizium anisopliae and Metarhizium marquandii species for the control of Nasutitermes sp. The strains of T. endophyticum (4.439), M. anisopliae (4.443) and M. marquandii (4.472) with their respective isolation codes, were evaluated using suspensions at concentrations of 105, 106, 107 and 108 conidia/mL against the termites. The fungi were characterized to species level by molecular analysis. The greatest virulence was registered with T. endophyticum (4.439), with a mortality of 100 % on the 4th day of treatment for all analyzed concentrations. The M. anisopliae strain (4.443) proved to be efficient, causing a mortality of 100 % on the 7th and 6th days at dilutions of 107 and 108 conidia/mL, respectively. Rates lower than 100 % were registered with M. marquandii (4.472). Therefore, the three fungal strains showed virulence against the termites Nasutitermes sp. In this study, the fungi Tolypocladium endophyticum and Metarhizium marquandii are reported for the first time for the biological control of pests, indicating the potential of native Amazonian fungi for the biological control of thermites Nasutitermes sp.

RESUMEN Las termitas del género Nasutitermes son consideradas una de las principales plagas urbanas y agroforestales de Brasil, donde el principal método de control es la aplicación de pesticidas. Los hongos entomopatógenos pueden ser usados como alternativa para reducir la población de esta plaga en el medio ambiente. Por tanto, el objetivo de este estudio fue evaluar la virulencia de aislados de hongos nativos de suelo amazónico pertenecientes a las especies Tolypocladium endophyticum, Metarhizium anisopliae y Metarhizium marquandii para el control de Nasutitermes sp. Las cepas de T. endophyticum (4,439), M. anisopliae (4,443) y M. marquandii (4,472) con sus respectivos códigos de aislamiento, fueron evaluadas utilizando suspensiones a concentraciones de 105, 106, 107 y 108 conidios/mL contra las termitas. Los hongos se caracterizaron mediante análisis moleculares para confirmar la especie. La mayor virulencia se registró con el hongo T. endophyticum (4,439), con una mortalidad del 100 % al cuarto día de tratamiento para todas las concentraciones analizadas. La cepa M. anisopliae (4,443) demostró ser eficiente, causando una mortalidad del 100 % al sexto y septimo días a las diluciones de 107 y 108 conidios / mL, respectivamente. Se registraron tasas inferiores al 100 % con M. marquandii (4,472). Por tanto, las tres cepas de hongos mostraron virulencia contra las termitas Nasutitermes sp. En este estudio, los hongos Tolypocladium endophyticum y Metarhizium marquandii son reportados por primera vez para el control biológico de plagas, indicando el potencial de hongos nativos de suelo amazónico para el control biológico de termitas Nasutitermes sp.

A Systematic Review on Bioactivity of Brazilian Mushrooms (Agaricomycetes).

The medicinal properties of Agaricomycetes mushrooms have been explored for millennia. Several biological activities produced by mushrooms have been evaluated worldwide. Therefore, this systematic review aims to present the studies that evaluated the biological activities demonstrated by mushrooms from Brazil. To select the articles, PubMed, Scopus, ScienceDirect and Google Scholar databases were used, with publications between the years 2001-2018, using the key words "mushrooms," "medicinal properties," "extracts," and "Brazil." The articles were selected according to three inclusion criteria: made with mushrooms from Brazil; title and summary with assessment of biological activity of basidioma or mycelium extracts; title and summary with evaluation of biological activity of isolated compound. Exclusion criteria were: work not done in Brazil; review articles; duplication of articles and abstracts and full texts irrelevant to the topics in question. A total of 31 articles were selected. According to the way used to assess biological activities, 54.8% evaluated from basidioma extract, 32.3% from isolated compound and 12.9% from mycelium extract. A total of 12 biological activities were evaluated, the most frequent were antioxidant (22%), anti-inflammatory (12.2%), anticancer (9.8%), antimicrobial (9.8%), and cytotoxic (7.3%). A total of 22 species of Agaricomycetes were evaluated in the studies, Agaricus blazei (=A. brasiliensis) the most studied (35.5%), mainly for antigenotoxic and anticlastogenic/antimutagenic activities. The most common isolated compounds of Agaricomycetes evaluated were ß-glucan, fucogalactane, mannogalactoglucan, amaurocin, and polysaccharides. Agaricomycetes studied in Brazil are promising for medicinal applications.

Study of Antibacterial Activity of Amazonian Agaricomycetes Mushrooms from Brazil.

This study aimed to examine the antibacterial activity of extracts of Amazonian Agaricomycetes macrofungi. The sample comprised 168 specimen collected at the Zoobotanical Park of the Federal University of Acre (Brazil), of which 32 (19%) were isolated. Extracts of the 32 isolates and 28 Agaricomycetes basidiomata were prepared and tested for activity against Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, and Klebsiella pneumoniae bacteria, using the disc diffusion method. Of the 32 metabolite extracts of the isolated Basidiomycetes, 11 (34.4%) exhibited antibacterial activity especially against Gram-negative E. coli and K. pneumoniae bacteria, while of the 28 basidiomata extracts, 6 (21.4%) exhibited antibacterial activity mainly against the Gram-positive S. aureus bacteria. This study is the first to report the antibacterial activity of the Agaricomycetes species Coriolopsis caperata, Cyclomyces iodinus, Cymatoderma sp., Favolus tenuiculus, and Tyromyces cf. polyporoides, and contributes as the first investigation about the antibacterial potential of Basidiomycetes grown in Southwest Amazonia.

Oral glutamine supplementation attenuates inflammation and oxidative stress-mediated skeletal muscle protein content degradation in immobilized rats: Role of 70 kDa heat shock protein.

Skeletal muscle disuse results in myofibrillar atrophy and protein degradation, via inflammatory and oxidative stress-mediated NF-kB signaling pathway activation. Nutritional interventions, such as l-glutamine (GLN) supplementation have shown antioxidant properties and cytoprotective effects through the modulation on the 70-kDa heat shock protein (HSP70) expression. However, these GLN-mediated effects on cell signaling pathways and biochemical mechanisms that control the myofibrillar protein content degradation in muscle disuse situations are poorly known yet. This study investigated the effects of oral GLN plus l-alanine (ALA; GLN â€‹+ â€‹ALA-solution) supplementation, either in their free or dipeptide (L-alanyl-l-glutamine-DIP) form, on GLN-glutathione (GSH) axis and cytoprotection mediated by HSP70 protein expression in the slow-twitch soleus and fast-twitch gastrocnemius skeletal muscle of rats submitted to 14-days of hindlimb immobilization-induced disuse muscle atrophy. Forty-eight Wistar rats were distributed into 6 groups: hindlimb immobilized (IMOB group) and hindlimb immobilized orally supplemented with either GLN (1 g kg-1) plus ALA (0.61 g kg-1) â€‹(GLN â€‹+ â€‹ALA-IMOB group) or 1.49 â€‹g â€‹kg-1 of DIP (DIP-IMOB group) and; no-immobilized (CTRL) and no-immobilized supplemented GLN â€‹+ â€‹ALA and DIP baselines groups. All animals, including CTRL and IMOB rats (water), were supplemented via intragastric gavage for 14 days, concomitantly to immobilization period. Plasma and muscle GLN levels, lipid (thiobarbituric acid reactive substances-TBARS) and protein (carbonyl) peroxidation, erythrocyte concentration of reduced GSH and GSH disulfide (GSSG), plasma and muscle pro-inflammatory TNF-α levels, muscle IKKα/ß-NF-kB signaling pathway and, the myofibrillar protein content (MPC) were measured. The MPC was significantly lower in IMOB rats, compared to CTRL, GLN â€‹+ â€‹ALA, and DIP animals (p â€‹< â€‹0.05). This finding was associated with reduced plasma and muscle GLN concentration, equally in IMOB animals. Conversely, both GLN â€‹+ â€‹ALA and DIP supplementation restored plasma and muscle GLN levels, which equilibrated GSH and intracellular redox status (GSSG/GSH ratio) in erythrocytes and skeletal muscle even as, increased muscle HSP70 protein expression; attenuating oxidative stress and TNF-α-mediated NF-kB pathway activation, fact that reverberated on reduction of MPC degradation in GLN â€‹+ â€‹ALA-IMOB and DIP-IMOB animals (p â€‹< â€‹0.05). In conclusion, the findings shown herein support the oral GLN â€‹+ â€‹ALA and DIP supplementations as a therapeutic and effective nutritional alternative to attenuate the deleterious effects of the skeletal muscle protein degradation induced by muscle disuse.

Antibacterial activity of fractions and isolates of Maytenus guianensis Klotzsch ex Reissek (Celastraceae) Chichuá Amazon.

INTRODUCTION: This aim of this study was to evaluate the antibacterial activity of fractions and isolates of Maytenus guianensis, a plant species used in Amazonian folk medicine. METHODS: A disk diffusion technique was used to investigate the antibacterial potential. RESULTS: The hexanic fractions and tingenone B isolate showed inhibitory effects against Staphylococcus aureus and Streptococcus pneumoniae. CONCLUSIONS: These results indicate the antibacterial potential of this species and will enable future studies to identify novel therapeutic alternatives from this species.

Antibacterial activity of fractions and isolates of Maytenus guianensis Klotzsch ex Reissek (Celastraceae) Chichuá Amazon

Abstract INTRODUCTION This aim of this study was to evaluate the antibacterial activity of fractions and isolates of Maytenus guianensis, a plant species used in Amazonian folk medicine. METHODS A disk diffusion technique was used to investigate the antibacterial potential. RESULTS The hexanic fractions and tingenone B isolate showed inhibitory effects against Staphylococcus aureus and Streptococcus pneumoniae. CONCLUSIONS These results indicate the antibacterial potential of this species and will enable future studies to identify novel therapeutic alternatives from this species.

Metabolites from endophytic Aspergillus fumigatus and their in vitro effect against the causal agent of tuberculosis / Metabolitos de Aspergillus fumigatus endofítico e seu efeito in vitro contra o agente causal da tuberculose

ABSTRACT Tuberculosis (TB) remains one of the most deadly communicable infectious diseases, causing 1.4 million deaths in 2015 worldwide due to many conditions, including the inadequate treatment and the emergence of multidrug-resistant strains of the causal agent, Mycobacterium tuberculosis. Therefore, drugs developed from natural sources, as microorganisms and plant extracts, are a frequent target for the research and discovery of antimicrobial compounds. The current study started the characterization of compounds produced by an Aspergillus fumigatus isolated from copaíba (Copaifera multijuga) that efficiently inhibits M. tuberculosis by releasing the compounds into the fermentation broth under specific culture conditions. A preliminary assay was carried out with a correlate species, M. smegmatis, aiming to detect an antimicrobial effect related to A. fumigatus fermentation broth. The direct use of this substrate in antibiosis assays againstM. tuberculosis H37Rv strain (ATCC 27294) allowed the detection of antimicrobial activity with a minimal inhibitory concentration of 256 μg mL-1, demonstrating that purification processes developed by the Biotage Flash Chromatography System are robust and reliable techniques for purification of compounds from natural sources. Also, this chromatographic system can be used in combination with specific biochemical tests, improving the search for reliable results. We conclude that this fraction can express a broad action range, inhibiting both Mycobacterium species used as target organisms.

RESUMO A tuberculose continua a ser uma das doenças infecciosas transmissíveis mais mortais, causando 1,4 milhão de mortes em 2015 em todo o mundo devido a vários fatores, incluindo o tratamento inadequado e o surgimento de cepas multirresistentes do agente causal, Mycobacterium tuberculosis. Portanto, as drogas desenvolvidas a partir de fontes naturais, como micro-organismos e extratos de plantas, são um alvo freqüente para a pesquisa e descoberta de compostos antimicrobianos. O presente estudo foi um ponto de partida para caracterizar compostos produzidos por um Aspergillus fumigatus isolado de copaíba (Copaifera multijuga) que inibe eficientemente M. tuberculosis, liberando os compostos no caldo de fermentação em condições de cultura específicas. Realizou-se um ensaio preliminar com uma espécie correlata, M. smegmatis, com o objetivo de detectar um efeito antimicrobiano relacionado ao caldo de fermentação de A. fumigatus. O uso direto deste substrato em ensaios de antibiose contra a estirpe H37Rv de M. tuberculosis (ATCC 27294) permitiu a detecção de atividade antimicrobiana com uma concentração inibitória mínima de 256 μg mL-1, demonstrando que os processos de purificação desenvolvidos pelo Biotage Flash Chromatography System são técnicas robustas e confiáveis para purificar compostos de fontes naturais. Além disso, este sistema cromatográfico pode ser usado em combinação com testes bioquímicos específicos, melhorando a busca de resultados confiáveis. Concluímos que esta fração pode expressar uma ampla gama de ação, inibindo ambas as espécies de Mycobacterium utilizadas como organismos-alvo.

Effects of culture filtrates of endophytic fungi obtained from Piper aduncum L. on the growth of mycobacterium tuberculosis

Substances that inhibit the growth of Mycobacterium tuberculosis could potentially be used as antibiotics. These substances could also be added to test culture media to improve the speed of tuberculosis diagnosis. The aim of this work was to investigate the influence of culture filtrates of endophytic fungi isolated from P. aduncum L. on the growth of M. tuberculosis. To achieve this objective, the following methodology was used: a) endophytic fungi were isolated from the leaves and stems of P. aduncum L.; b) the isolated fungi were submitted to submerged bioprocessing; c) culture filtrates from the bioprocess were assayed to evaluate their effect on the growth of M. tuberculosis. We isolated 315 fungal types, which represented 85 morphologies, from different parts of P. aduncum L. The bioassays were performed on 82 culture filtrates and 6 plant extracts and resulted in the detection of 1 culture filtrate that stimulated the growth of M. tuberculosis and 15 that inhibited microbial growth. None of the phytochemical extracts had an effect on the growth of M. tuberculosis. In conclusion, we observed that the endophytic fungi isolated from P. aduncum L. (Piperaceae) produced extracellular metabolites (present in the culture filtrate) that affect the growth of M. tuberculosis. These compounds have the potential to be used as antimicrobials or in the diagnosis of tuberculosis.

PKO: alternative method for isolating mycobacteria from sputum

We elaborated an alternative culture method, which we denominated PKO (initials in tribute of respect to Petroff, Kudoh and Ogawa), for isolating Mycobacterium tuberculosis from sputum for diagnosis of pulmonary tuberculosis (TB), and to compare its performance with the Swab and Petroff methods. For the technique validation, sputum samples from patients suspected of pulmonary TB cases were examined by acid-fast microscopy (direct and concentrated smear), PKO, Swab and Petroff methods. We found that Petroff and PKO methods have parity in the effectiveness of M. tuberculosis isolation. However, by the PKO method, 65 percent of isolated strains were detected in a period of £15 days, while by the Petroff method the best detection was in an interval of 16-29 days (71 percent). In positive smear samples, the average time of PKO isolation is only superior to the one related for Bactec 460TB. In conclusion, the exclusion of the neutralization stage of pH in the PKO reduces the manipulation of the samples, diminishes the execution time of the culture according to the Petroff method and facilitates the qualification of professionals involved in the laboratorial diagnosis of Tuberculosis.

Foi elaborado um método de cultivo alternativo, denominado por nós PKO (iniciais referentes à Petroff, Kudoh e Ogawa), para o isolamento do Mycobacterium tuberculosis em amostras de escarro para o diagnóstico da tuberculose pulmonar (TB). Para validação da técnica, amostras de escarro de pacientes suspeitos de TB foram submetidas aos métodos de baciloscopia (direta e pós-concentração), PKO, Swab e Petroff. A análise comparativa entre o método de Petroff e o PKO mostrou paridade de resultados em relação ao isolamento e número de colônias de M. tuberculosis. Porém, pelo método PKO, 65 por cento das cepas isoladas foi detectada em um período £15 dias, enquanto que pelo método de Petroff a melhor detecção ocorreu em um intervalo de 16-29 dias (71 por cento). O tempo médio de isolamento pelo PKO é somente superior ao sistema comercial Bactec 460TB em amostras positivas na baciloscopia. A exclusão da etapa de neutralização de pH no método PKO reduz a manipulação das amostras, diminui o tempo de execução do cultivo em relação ao de Petroff e facilita o treinamento de profissionais que realizam o diagnóstico laboratorial da TB.