RESUMO: Actinobacillus pleuropneumoniae é o agente causador da pleuropneumonia suína, uma doença respiratória contagiosa associada à alta morbidade e perdas econômicas. Embora a antibioticoterapia ajude a controlar a disseminação do patógeno na fazenda, a resistência a várias classes de antibióticos tem sido relatada e o tratamento pode ser prejudicado pela capacidade bacteriana de formar biofilmes. Isso aumenta a necessidade de abordagens terapêuticas alternativas, que incluem o uso de compostos naturais com atividades antimicrobianas e/ou antibiofilme. Neste trabalho, analisamos, pelo método de microdiluição em caldo nutriente, os efeitos inibitórios e bactericidas dos óleos essenciais obtidos de dezoito plantas medicinais brasileiras populares e/ou temperos contra isolados clínicos de Actinobacillus pleuropneumoniae. Então, concentrações subinibitórias dos óleos ativos foram testados quanto a suas atividades antibiofilme, analisados pelo método do cristal violeta. Dentre os dezoito óleos testados, oito (extraídos da canela, coentro, hortelã-pimenta, hortelã, tomilho, manjerona, eucalipto e louro) apresentaram atividade bacteriostática e bactericida contra todos os isolados, e concentrações subinibitórias de cinco deles romperam biofilmes pré-formados em até 80%, além de inibirem fortemente a formação de biofilmes. A composição química desses óleos foi avaliada por cromatografia gasosa/espectrometria de massa (CG/MS) e é apresentada, indicando que suas propriedades bactericidas e antibiofilme estavam principalmente associadas à presença de monoterpenos e fenilpropanóides. Este é o primeiro relato de óleos essenciais com potencial para controlar a contaminação ambiental e infecção animal por A. pleuropneumoniae, representando uma alternativa contra o crescente aumento de resistência bacteriana aos antibióticos.

ABSTRACT: Actinobacilluspleuropneumoniae is the causative agent of swine pleuropneumonia, a contagious respiratory disease associated with high morbidity and economic losses. While antibiotic therapy helps to control the spreading of the pathogen on the farm, resistance to several classes of antibiotics were reported, and treatment can be impaired by the bacterial ability to form biofilms. This increases the need for alternative therapy approaches, including the use of natural compounds with antimicrobial and/or antibiofilm activities. In this research we analyzed, by the broth microdilution method, the inhibitory and bactericidal activities of the essential oils obtained from eighteen Brazilian popular medicinal plants or spices against clinical isolates of Actinobacilluspleuropneumoniae. After that, sub-inhibitory concentrations of active oils were tested for their antibiofilm effects, analyzed by the crystal violet method. Among the eighteen oils tested, eight (extracted from cinnamon, coriander, peppermint, spearmint, thyme, marjoram, eucalyptus, and laurel) presented bacteriostatic and bactericidal activities against all isolates, and subinhibitory concentrations of five of them disrupted up to 80% preformed biofilms, and significantly inhibited biofilm formation. The chemical composition of such oils was assessed by gas chromatography/mass spectrometry (GC/MS) and is presented, indicating that their bactericidal and antibiofilm properties were mostly associated with the presence of monoterpenes and phenylpropanoids. To our knowledge, this is the first report of essential oils with potential to control environmental contamination and animal infection with A. pleuropneumoniae, representing an alternative to increasing levels of antibiotic resistance.

Abstract Adenocalymma axillarum (K.Schum.) L.G. Lohmann is a liana belonging to the family Bignoniaceae. In traditional medicine, the genus Adenocalymma is used to treat fever, skin ailments, and body, joint, and facial muscle pains, and it is also applied as cosmetic. Biological assays conducted with the A. axillarum crude leaf ethanol extract have indicated leishmanicidal activity and absence of cytotoxicity. This study aimed to analyze the A. axillarum leaf ethanol crude extract by high-performance liquid chromatography-high-resolution mass spectrometry- diode array detector (HPLC-HRMS-DAD) and to evaluate the leishmanicidal and cytotoxic activities of this crude extract, its fractions, and isolated compounds. HPLC-HRMS-DAD analysis of this extract revealed that it consisted mainly of flavonoids, with nine major compounds. Extract purification yielded 4-hydroxy-N-methylproline, 6-β-hydroxyipolamiide, quercetin-3-O-robinobioside, hyperin, isorhamnetin-3-O-robinobioside, and 3’-O-methylhyperin, which were identified by Nuclear Magnetic Resonance. The isolated compounds were inactive against Leishmania amazonensis promastigotes and human lung fibroblast cells.

ABSTRACT An adequate combination of factors involved in the technology used for phytosanitary product application contributes to an efficient spray deposition on the target. The objective of this study was to use multivariate analysis to characterize the magnitude of effects and the order of influence of three factors that interfere with the quality of phytosanitary product application in coffee plants. An entirely randomized design was adopted, with four repetitions, using a 2 × 2 × 3 factorial scheme, with two classes of droplets quality (fine and coarse), two application rates (250 and 400 L ha-1), and the use of adjuvants (with no adjuvant or with Fighter®and Aureo® adjuvants). The quality of the application was determined by jointly analyzing the spray deposition on three thirds of leaves, in their internal and external layers, the runoff to soil, coverage, droplet density, relative amplitude, and the volumetric median diameter. The results underwent analysis of variance (ANOVA) to measure the effect sizes (η2). After testing the assumptions of multivariate analysis, clustering and principal component analyses were performed. The class of droplets was found to be the most influential factor in the quality of the phytosanitary product application (spray deposition and runoff to soil). When focusing on spray deposition on leaves, the second-most influential factor was the application rate and the relation between the application rate and the adjuvants. For the other variables, the second-most influential factor was the application rate.

Fridericia speciosa (Bignoniaceae) is a liana that occurs in humid to dry vegetation in eastern and southeastern Brazil. Species belonging to the genus Fridericia are employed as medicinal plants. This study evaluates the antipromastigote and cytotoxic activities of the crude extract, fractions, and compounds isolated from F. speciosa, and an HPLC (High-Performance Liquid Chromatography) analytical method was used to quantify the bioactive flavonoids. The flavonoids alpinetin (1), luteolin (2), diosmetin (3), and apigenin (4) were isolated from the active EtOAc-soluble fraction (EAF) of the crude ethanol extract (LCE) obtained from F. speciosa leaves. Both LCE and EAF were active against the normal human lung fibroblast cell line GM07492A (IC50 of 460.7 µg mL-1 and 381.4 µg mL-1, respectively). Only EAF showed activity against Leishmania amazonensis (80.4% at 50 µg mL-1). Compounds 1 and 2 were quantified by HPLC-DAD in both LCE (1: 6.21%; 2: 1.11%) and EAF (1: 45.32%; 2: 2.00%). At 100 µM, compounds1-4 inhibited L. amazonensis flagellar motility by 40.2%, 26.8%, 1.0%, and 24.8%, respectively, while their toxicities (IC50) to the normal cell were > 1000, 353.0, 766.2, and >1000 µg mL-1, respectively. Therefore, F. speciosa leaves are a prolific source of bioactive flavonoids.

The present work studied aqueous extracts of castor beans as a natural inhibitor of the corrosion of mild steel in 1 M HCl using gravimetric tests, potentiodynamic polarization curves, linear polarization method, electrochemical impedance measurements, surface analysis by scanning electron microscopy (SEM) and chemical characterization by Fourier transform infrared spectroscopy (FTIR). Gravimetric tests showed that inhibition efficiency (IE) increased with immersion time and concentration of the extract, reaching 97.8% in the presence of 800 mg L−1 inhibitor after 24 h immersion time. For the gravimetric tests varying the temperature, a decrease of the activation energy (Ea) was observed, suggesting chemical adsorption of extract components on the metal surface. The extracts from the castor beans acted as a good corrosion inhibitor, and the macromolecules present in the high molecular weight fraction (HMWF) have a fundamental participation in this process since it presented 93.7% of IE for 200 mg L−1.

Lipoxygenase (LOX) plays an important role in inflammatory processes and Arrabidaea brachypoda (DC.) Bureau (Bignoniaceae) has been described as presenting anti-inflammatory activity. Therefore, the objective of this study was to develop a high-performance liquid chromatography (HPLC) procedure to directly recognize LOX inhibitor compounds in A. brachypoda crude extract, facilitating the isolation, characterization of bioactive compounds, evaluation of natural compounds using an in vitro 15-LOX assay and prediction of the most probable binding modes of their main constituent through molecular docking simulations. The chemical analysis was performed by ethanol crude extract microfractionation using HPLC-DAD (diode array detector) associated to a fraction collector. The bioactive chromatogram displayed a peak with 50.9% LOX inhibition at 13.6 min retention time. The extract was purified and conandroside was isolated, presenting a LOX inhibitory activity with an inhibitory concentration (IC50) of 7.8 ± 1.1 µM, close to standard quercetin (IC50 7.6 ± 0.3 µM). Additionally, conandroside was not cytotoxic to normal cells (GM07492A). The LOX-conandroside complex displayed a slightly higher docking score (92.7) than quercetin (71.5). These results together suggest that conandroside could be explored as lipoxygenase inhibitor.

RESUMO A introdução de espécies exóticas é considerada uma das principais causas da perda de biodiversidade. A Floresta Nacional de Carajás é uma das maiores províncias minerais do mundo. As atividades da mineração provocaram alterações dos habitats naturais, deixando áreas degradadas após a exploração mineral. Uma das áreas de mineração dentro da FLONA de Carajás foi usada para a extração de ouro. No processo de exploração, uma enorme cava foi formada pela remoção do solo que foi colocado em montes nas proximidades. Para evitar a erosão do solo desses montes, uma gramínea exótica, Urochloa decumbens (Stapf) R.D.Webster (Poaceae) foi plantada. O objetivo deste estudo foi comparar o impacto desta gramínea não-nativa na riqueza de espécies e composição de espécies da regeneração natural nas áreas degradadas. Quatro áreas foram comparadas, duas com e duas sem a presença de U. decumbens. Em cada área, vinte e quatro parcelas de 1m² foram estabelecidas. A riqueza de espécies e os tamanhos das populações da regeneração natural foram significativamente menores nas parcelas onde a gramínea exótica estava presente. Nosso estudo mostra que U. decumbens tem um efeito negativo sobre a riqueza de espécies e densidade populacional e sua presença alterou a composição e distribuição de formas de vida da regeneração natural das espécies.

ABSTRACT The introduction of exotic species is considered as one of the major causes of biodiversity loss. The National Forest of Carajás is one of the largest mineral provinces in the world. Mining activities caused changes of the natural habitats, leaving degraded areas after the mineral exploitation. One of the mining areas within FLONA Carajás was used for the extraction of gold. In the process of exploitation, a huge depression was formed by the removal of soil which was mounded up nearby. To prevent soil erosion of these mounds, an exotic grass, Urochloa decumbens (Stapf) R.D.Webster (Poaceae) was planted. The objective of this study was to compare the impact of this non-native grass on species richness and species composition of the natural regeneration in the degraded areas. Four areas were compared, two with and two without presence of U. decumbens. In each area, twenty four 1m²/plots were established. Species richness of the regeneration areas and population sizes were significantly lower in the plots where the exotic grass was present. Our study shows that U. decumbens had a negative effect on species richness and population density, and its presence changed the species composition and distribution of life forms of the natural regeneration.

Os principais rios da bacia Amazônia e sua biota estão ameaçados pelos planos de construção de grandes usinas hidrelétricas que poderá causar fortes impactos nas comunidades vegetais das áreas alagadas. O objetivo desse estudo é apresentar os inventários botânicos em três áreas de vegetações inundadas dos rios Tocantins, Xingu e Tapajós no leste da Amazônia. Os resultados do estudo mostram uma flora altamente especializada das vegetações inundadas, claramente distintas entre as três bacias, embora os ambientes entre as bacias dos rios não sejam distintos entre si e as condições ambientais muito semelhantes. Somente 6 das 74 espécies ocorreram nos três inventários, sendo a maioria das arvores e arbustos restritos a um dos rios, indicando um alto grau de distribuição local. Espécies diferentes ocupam nichos ambientais semelhantes, mostrando alto valor de conservação dessas formações vegetais. Os planos de conservação devem considerar a complementaridade das espécies nas tomadas de decisões de onde colocar as unidades de conservação nesses nas florestas inundadas a fim de evitar de forma irreversível a perda da biodiversidade.

The major rivers of the Amazon River basin and their biota are threatened by the planned construction of large hydroelectric dams that are expected to have strong impacts on floodplain plant communities. The present study presents forest inventories from three floodplain sites colonized by alluvial riparian vegetation in the Tapajós, Xingu and Tocantins River basins in eastern Amazonian. Results indicate that tree species of the highly specialized alluvial riparian vegetation are clearly distinct among the three river basins, although they are not very distinct from each other and environmental constraints are very similar. With only 6 of 74 species occurring in all three inventories, most tree and shrub species are restricted to only one of the rivers, indicating a high degree of local distribution. Different species occupy similar environmental niches, making these fragile riparian formations highly valuable. Conservation plans must consider species complementarily when decisions are made on where to place floodplain forest conservation units to avoid the irreversible loss of unique alluvial riparian vegetation biodiversity.

A simple flow system with multiple pulse amperometric detection using a single working electrode is proposed for simultaneous determination of ascorbic (AA) and acetylsalicylic (AAS) acids in pharmaceutical formulations. The procedure is based on application of two potential pulses: 0.90 V/50 ms: oxidation and determination of AA without the interference of AAS; 1.35 V/50 ms: oxidation of both compounds and quantification of AAS by current subtraction using a correction factor. Sampling rate was estimated as 125 injections per hour and the limits of detection were 0.17 and 0.16 µmol L-1 for AA and AAS, respectively. Results for commercial samples agreed with those obtained using HPLC.

This study developed a predictive model to identify pleural tuberculosis. A consecutive cases study of patients investigating the cause of pleural effusion, in an area of high prevalence of tuberculosis (Rio de Janeiro, Brazil). Clinical and laboratory variables were compared among patients with tuberculosis (TB) and without tuberculosis (NTB), individually and using logistic regression. The performance was described as diagnostic accuracy, compared to a gold standard in a masked way. We have studied 104 TB patients, 41 with malignant, 29 transudates, 28 parapneumonic, 13 with miscellaneous diseases. After identification of individual discrimination power aided by clinical, radiological and laboratory variables, the following ones were included in a multivariate analysis: ADA, total leukocytes, percentile of lymphocytes, protein, lactate dehydrogenase, duration of disease, age and gender. A logistic regression model to predict pleural tuberculosis including the five first variables showed the best performance. A receiver operating characteristic curve identified the best cutoff at 0.7, resulting in a sensitivity and specificity of more then 95%. The predictive model improved the specificity of ADA alone, keeping its sensitivity. This model seems helpful when a microbiological or histological diagnosis of pleural tuberculosis could not be established. External validation of these results is necessary before recommendation for routine application.

Azadirachta indica A. Juss. é uma espécie nativa das regiões áridas da Ásia e África, mas que atualmente encontra-se distribuída também nas Américas e outras regiões de clima tropical e sub-tropical. É uma planta muito resistente, que apresenta excelentes propriedades medicinais. O objetivo deste trabalho foi definir o tipo de substrato e a temperatura mais adequada à germinação de sementes de nim-indiano (Azadirachta indica A. Juss.) e conhecer a morfologia de seu desenvolvimento pós-seminal, caracterizando as plântulas normais e o tipo de germinação. As sementes foram semeadas nos substratos vermiculita, areia e rolo de papel germitest e mantidas em germinador nas temperaturas de 25°C, 30°C, 35°C e 20-30°C. O delineamento estatístico utilizado foi o inteiramente casualizado, com quatro repetições de 25 sementes, avaliando-se a porcentagem de germinação. Os melhores resultados da germinação são verificados na temperatura constante de 35°C, nos substratos vermiculita e em rolo de papel com contagens de 15 e 30 dias. As plântulas normais são caracterizadas como de germinação epígea.

Azadirachta indica A. Juss. is a native specie of the arid areas of Asia and Africa, but nowadays is also distributed in America and other areas with tropical and sub-tropical climate. It is a very resistant plant that presents excellent medicinal properties. The objective of this study was to define the ideal germination conditions for neem (Azadirachta indica A. Juss.) seeds evaluating the most appropriate substrates, temperature and characterizing its post-seminal morphology. Seeds were sown on vermiculite, sand and towel paper and maintained in a germinator at 25°C, 30°C, 35°C and 20-30°C. A complete randomized design was used, with 25 seeds sown in four replications, and the germination percentage was evaluated. The best results were obtained for temperature of 35°C in vermiculite and towel paper substrates and countings performed at 15 and 30 days after sowing. Normal seedlings were classified as epigeal germination.