Revisiting molecular diagnostics of Streptococcus pneumoniae

Streptococcus pneumoniae is a human pathobiont that colonizes the nasopharynx. S. pneumoniae is responsible for causing non-invasive and invasive disease such as otitis, pneumonia, meningitis, and sepsis, being a leading cause of infectious diseases worldwide. Due to similarities with closely related species sharing the same niche, it may be a challenge to correctly distinguish S. pneumoniae from its relatives when using only non-culture based methods such as real time PCR (qPCR). In 2007, a molecular method targeting the major autolysin (lytA) of S. pneumoniae by a qPCR assay was proposed by Carvalho and collaborators to identify pneumococcus. Since then, this method has been widely used worldwide. In 2013, the gene encoding for the ABC iron transporter lipoprotein PiaA, was proposed by Trzcinzki and collaborators to be used in parallel with the lytA qPCR assay. However, the presence of lytA gene homologues has been described in closely related species such as S. pseudopneumoniae and S. mitis and the presence of piaA gene is not ubiquitous between S. pneumoniae. The hyaluronate lyase gene (hylA) has been described to be ubiquitous in S. pneumoniae. This gene has not been used so far as a target for the identification of S. pneumoniae. The aims of our study were to evaluate the specificity, sensitivity, positive predicted value (PPV) and negative predicted value (NPV) of the lytA and piaA qPCR methods; design and implement a new assay targeting the hylA gene and evaluate the same parameters above described; analyze the assays independently and the possible combinations to access what is the best approach using qPCR to identify S. pneumoniae. A total of 278 previously characterized strains were tested: 61 S. pseudopneumoniae, 37 Viridans group strains, 30 type strains from other streptococcal species and 150 S. pneumoniae strains. The collection included both carriage and disease isolates. By Mulilocus Sequence Analysis (MLSA) we confirmed that strains of S. pseudopneumoniae could be misidentified as S. pneumoniae when lytA qPCR assay is used. The results showed that as a single target, lytA had the best combination of specificity, sensitivity, PPV and NPV being, 98.5%, 100.0%, 98.7% and 100.0% respectively. The combination of targets with the best values of specificity, sensibility, PPV and NPV were lytA and piaA, with 100.0%, 93.3%, 97.9% and 92.6%, respectively. Nonetheless by MLSA we confirmed that strains of S. pseudopneumoniae could be misidentified as S. pneumoniae and some capsulated (23F, 6B and 11A) and non-capsulated S. pneumoniae were not Identified using this assay. The hylA gene as a single target had the lowest PPV. Nonetheless it was capable to correctly identify all S. pneumoniae.

UV-Vis, FTIR and antioxidant study of Persea Americana (Avocado) leaf and fruit: A comparison

El aguacate (Persea americana Miller) es una conocida fruta arb´orea con un alto contenido nutricional que crece en varias partes del mundo. El presente estudio compara los espectros del UV-Vis y del espectr´ometro infrarrojo con transformada de Fourier (FTIR) de la fruta y de la hoja de aguacate (c´ascara, pulpa y aceite) cultivado en Ecuador y posteriormente eval´ua su actividad antioxidante empleando el 1,1-difenil-2-picrilhidrazil (DPPH•). El estudio de los espctros UV-Vis y FTIR revel´o que el aguacate tiene predominantemente flavonoides. Entre la hoja y el fruto del aguacate, se comprob´o mediante el ensayo DPPH• (captura de radicales libres), que la hoja tuvo una mayor actividad antioxidante, la que oscila entre 84,46% y 80,12%, con valores de 32.60-32.73 μg equivalentes de ´acido g´alico por mL. Se demostr´o que el orden de la actividad antioxidante de los extractos es: hoja de aguacate > c´ascara > aceite > pulpa. La actividad antioxidante tuvo una correlaci´on positiva con el contenido total de flavonoides y estos extractos de plantas (especialmente de las hojas del aguacate) son ´utiles para el desarrollo de futuros productos antioxidantes.

Atividade antimalárica de plantas medicinais da biorregião do Araripe-CE em modelo murino - Plasmodium berghei

Malaria, also popularly known as maleita , intermittent fever, paludism, impaludism, third fever or fourth fever, is an acute infectious febrile disease, which, in human beings, is caused by four species: Plasmodium falciparum, P. vivax, P. malariae and P. ovale. Malaria, one of the main infectious diseases in the world, is the most important parasitoses, with 250 million annual cases and more than 1 million deaths per year, mainly in children younger than live years of age. The prophylactic and therapeutic arsenal against malaria is quite restricted, since all the antimalarials currently in use have some limitation. Many plant species belonging to several families have been tested in vivo, using the murine experimental model Plasmodium berghei or in vitro against P. falciparum, and this search has been directed toward plants with antithermal, antimalarial or antiinflammatory properties used in popular Brazilian bolk medicine. Studies assessing the biological activity of medicinal plant essential oils have revealed activities of interest, such as insecticidal, spasmolytic and antiplasmodic action. It has also been scientifically established that around 60% of essential oils have antifungal properties and that 35% exhibit antibacterial properties. In our investigation, essential oils were obtained from the species Vanillosmopsis arborea, Lippia sidoides and Croton zethneri which are found in the bioregion of Araripe-Ceará. The chemical composition of these essential oils was partially characterized and the presence of monoterpenes and sesquiterpenes. The acute toxicity of these oils was assessed in healthy mice at different doses applied on a single day and on four consecutive days, and in vitro cytotoxicity in HeLa and Raw cell lines was determined at different concentrations. The in vivo tests obtained lethal dose values of 7,1 mg/Kg (doses administered on a single day) and 1,8 mg/Kg (doses administered over four days) for 50% of the animals. In the in vitro tests, the inhibitory concentration for 50% of cell growth in Hela cell lines was 588 μg/mL (essential oil from C. zethneri after 48 h), from 340-555 μg/mL (essential oil from L. sidoides, after 24 and 48 h). The essential oil from V. arborea showed no cytotoxicity and none of the essential oils were cytotoxic in Raw cell lines. These data suggest a moderate toxicity in the essential XVIII oils under study, a finding that does not impede their testing in in vivo antimalarial assays. Was shown the antimalarial activity of the essential oils in mice infected with P. berghei was assessed. The three species showed antimalarial activity from 36%-57% for the essential oil from the stem of V. arborea; from 32%-82% for the essential oil from the leaves of L. sidoides and from 40%-70% of reduction for the essential oil from the leaves of C. zethneri. This is the first study showing evidence of antimalarial activity with these species from northeast Brazil. Further studies to isolate the active ingredients of these oils are needed to determine if a single active ingredient accounts for the antimalarial activity or if a complex integration of all the compounds present occurs, a situation reflected in their biological activity

Anelagem de árvores como tratamento silvicultural em florestas naturais da Amazônia brasileira.

São avaliadas a eficácia de dois tipos de anelagem de árvores, como tratamento silvicultural para florestas naturais da Amazônia e a resistência de oito espécies arbóreas ao tratamento de anelagem, em 5 ha, na Floresta Nacional do Tapajós, à margem da BR 163, km 69, no município de Belterra, Pará. As espécies escolhidas foram: Carapa guianensis (andiroba, Meliaceae); Pouteria heterosepala (abiu, Sapotaceae); Hevea guianensis (seringueira, Euphorbiaceae); Helicostylis pedunculata (muiratinga-folha-peluda, Moraceae); Couratari oblongtfolia (tauari, Lecythidaceae); Sclerolobium crysophyllum (taxi-vermelho, Leguminosae); Virola melinonii (ucuuba-da-terra-firme, Myristicaceae); Bixa arborea (urucu-da-mata, Bixaceae). Os dois tipos de anelagem: anelagem completa e anelagem com entalhes foram analisados em três classes diamétricas, perfazendo seis tratamentos. De cada espécie foram escolhidas duas árvores, perfazendo um total de dezesseis árvores em cada tratamento. Tanto a anelagem completa quanto a anelagem com entalhes podem ser considerados tratamentos eficazes quando utilizado em desbastes, pois já aos cinco anos mostraram uma taxa de mortalidade alta. A anelagem completa se mostrou mais indicada como tratamento silvicultural, porque mostra a maior taxa de mortalidade em todos os tratamentos e dificulta a recuperação da área anelada. As espécies Pouteria heterosepala, Helicostylis pedunculata, Hevea guianensis e Couratari oblongifolia se mostraram mais resistentes a ambos os tipos de anelagem, com uma taxa de mortalidade considerada baixa. Nas espécies Sclerolobium crysophyllum, Virola melinonii, Bixa arborea e Garapa guianensis, a anelagem pode ser aplicada com sucesso, considerando que aos cinco anos já atingiram uma taxa média consideravelmente alta (maior que 60%), sem a utilização de arboricidas.

Changes in the spatial distribution of tree species in a terra firme rain forest in Brazilian Amazonia after logging.

The objective of this paper was to determine changes in the spatial distribution of tree species in a logged compared to an unlogged forest of the Tapajos National Forest in the municipality of Belterra, State of Para, Brazil, over an eight-year period. The distribution pattern was determined for trees> 5 cm dbh and, also, for trees > 30 cm dbh. The relationship (a quadrate method) discussed by McGinnis was selected to be used in this study. Forty-seven percent of species with trees > 5 cm dbh showed clumped distribution in the studied forests. Geissospermwn sericeunz Benth & Hook., Minquartia guianensis Aubl., Poureria bilocularis (H. Winkler) Bachni, Protium guacayantan Cuatrec, Sclerolobium chrysophyllunz Poepp. et Endl. and the Sapotaceae family (9 species) occurred in clumps of small trees (5 cm 5 dbh < 30 cm) and big trees (dbh > 30 cm) in both the logged and undisturbed forest. Trees in all sizes of these species certainly have aggregation characteristics in different light condition's during the whole growth-cycle. Only Sclerolobium cizzysophylltan out of fourteen species that occurred aggregated in all forest conditions was light demanding. The shade-tolerant Lecythis lurida (Miers) Mori and Manilkara huberi (Ducke) Stand!. showed also aggregated distribution for small and big trees in the unlogged forest. An aggregated distribution is not always directly correlated to abundance, considering that most of the clumped species had less than seven trees per hectare.

Restauração de florestas exploradas seletivamente no Acre.

Por meio de técnicas de restauração, florestas exauridas podem ser conduzidas de maneira a minimizar os efeitos da exploração seletiva que as modificaram. O plantio de mudas é um método rápido e eficaz de restauração de florestas. Este trabalho objetiva descrever os resultados de plantios de enriquecimento de florestas de produção nos municípios de Xapuri, Brasiléia e Rio Branco, no estado do Acre. Foram utilizadas dez espécies florestais madeireiras comerciais: amarelão (Aspidosperma vargasii A. DC.), angelim (Ormosia arborea (Vell.) Harms), cedro (Cedrela odorata L.), cerejeira (Amburana acreana (Ducke) A. C. Sm.), freijó (Cordia alliodora (Ruiz & Pav.) Oken), ipê (Tabebuia serratifolia (Vahl) G. Nicholson), itaúba (Mezilaurus itauba (Meisn.) Taub. ex Mez), jatobá (Hymenaea courbaril L.), mogno (Swietenia macrophylla King) e timbaúba (Enterolobium maximum Ducke). Os plantios foram entre outubro/2011 a março/2012, totalizando 1273 mudas. Os tratamentos silviculturais de condução e o monitoramento foram nos anos de 2012, 2013, 2014 e 2015. Após 48 meses ao plantio, a taxa média de sobrevivência foi de 42,3%, altura total média de 1,52 m e diâmetro médio de 1,88 cm. As espécies com os melhores desempenhos com relação a crescimento e sobrevivência foram cerejeira, cedro, freijó, jatobá, mogno e timbaúba.