ABSTRACT Paracoccidioides species cause paracoccidioidomycosis (PCM), a systemic mycosis highly prevalent in Brazil. Therapy of PCM has some issues that make studies for new therapeutic and vaccine targets relevant, such as the P. brasiliensis 60-kDa-heat-shock protein (PbHsp60), an immunogenic antigen that induces protection in experimental mice infection. Here, we investigated the relative expression of mRNA for PbHsp60 in P. brasiliensis in the different morphotypes of P. brasiliensis and in morphological transition phases. In addition, antibodies to rPbHsp60 were produced and used to analyze the location of PbHsp60 in yeast and hyphae by electron microscopy. The analyses showed a substantial increase in the relative amounts of HSP60 mRNA in yeast when compared to mycelium and an intermediate expression in transitional forms. Regarding the cell location, immunoelectron microscopy analysis revealed that PbHsp60 is within the cell wall. These observations suggest that this protein may be involved in the maintenance of the cell wall integrity and the interaction with the host for colonization, infection and pathogenesis.

In DNA vaccines, the gene of interest is cloned into a bacterial plasmid that is engineered to induce protein production for long periods in eukaryotic cells. Previous research has shown that the intramuscular immunization of BALB/c mice with a naked plasmid DNA fragment encoding the Mycobacterium leprae 65-kDa heat-shock protein (pcDNA3-Hsp65) induces protection against M. tuberculosis challenge. A key stage in the protective immune response after immunization is the generation of memory T cells. Previously, we have shown that B cells capture plasmid DNA-Hsp65 and thereby modulate the formation of CD8+ memory T cells after M. tuberculosis challenge in mice. Therefore, clarifying how B cells act as part of the protective immune response after DNA immunization is important for the development of more-effective vaccines. The aim of this study was to investigate the mechanisms by which B cells modulate memory T cells after DNA-Hsp65 immunization. C57BL/6 and BKO mice were injected three times, at 15-day intervals, with 100 µg naked pcDNA-Hsp65 per mouse. Thirty days after immunization, the percentages of effector memory T (TEM) cells (CD4+ and CD8+/CD44high/CD62Llow) and memory CD8+ T cells (CD8+/CD44high/CD62Llow/CD127+) were measured with flow cytometry. Interferon γ, interleukin 12 (IL-12), and IL-10 mRNAs were also quantified in whole spleen cells and purified B cells (CD43−) with real-time qPCR. Our data suggest that a B-cell subpopulation expressing IL-10 downregulated proinflammatory cytokine expression in the spleen, increasing the survival of CD4+ TEM cells and CD8+ TEM/CD127+ cells.

Here we describe the total syntheses and characterization by elemental analyses, infrared and NMR spectroscopy of three new compounds analogous to avenaciolide, a bis-γ-lactone isolated from Aspergillus avenaceus that possesses antifungal activity, where the octyl group of the natural product was replaced by aromatic groups containing chlorine and fluorine atoms. The effects of the avenaciolide, the novel compounds and their synthetic precursors on mycelia development and conidia germination of Colletotrichum gloeosporioides and Fusarium solani were evaluated in vitro. The title compounds were almost as active as avenaciolide. The absolute structures of the chlorinated analogs were determined by X-ray diffraction analysis.

Avaliaram-se as alterações nos indicadores do solo pH em água, MO e N totais, catiões de troca, P, K, Cu e Zn “assimiláveis”, metais pesados (Cd, Cr, Cu, Ni, Pb e Zn) e bactérias de origem fecal, entre 2004 e 2007, em dois campos experimentais com pastagens, por efeito da aplicação de lama residual urbana (LRU) e da fertilização mineral. As modalidades incluíram as combinações de dois tipos de pastagem (natural e semeada) com três tratamentos (0, LRU e Adubação), na Herdade do Revez (Luvissolos e Leptossolos Háplicos) e com quatro tratamentos (0, LRU, LRU+Calagem e Adubação+Calagem), na Herdade da Criméia (Luvissolos Férricos). Foram os teores médios de MO total e P “assimilável” as características químicas, comuns aos solos dos dois campos, em que o efeito benéfico das LRU na camada superficial foi significativo (P ≤ 5%). A aplicação de LRU também induziu os maiores acréscimos significativos de N total e de Mg de troca no solo da Herdade do Revez. A calagem elevou o pH e o teor de Mg de troca do solo, na Herdade da Criméia. As LRU não contaminaram os solos com metais pesados. Os níveis populacionais de coliformes e de enterococos fecais aumentaram, logo após a aplicação das LRU, para decrescerem progressivamente ao longo do tempo, igualando-se aos das testemunhas.

We evaluated the changes in the soil indicators pH in water, total OM and N, exchangeable cations, available P, K, Cu, and Zn, heavy metals (Cd, Cr, Cu, Ni, Pb, and Zn), and faecal origin bacteria, between 2004 and 2007, as affected by sewage sludge (SS) application and mineral fertilisation in two field experiments with pastures. The treatments were the combinations of two types of pasture (natural and sown) with three fertilisation treatments (0, SS, inorganic fertilisers) in the experiment of Herdade do Revez (Haplic Luvisols and Haplic Leptosols), and four fertilisation treatments (0, SS, Liming+SS, and Liming+inorganic fertilisers) in Herdade da Criméia (Ferric Luvisols). The OM and available P contents were the chemical properties that significantly increased (P ≤ 5%) in the soil surface layer of both field experiments. The SS application also caused the highest significant increases in soil total N and exchangeable Mg contents in Herdade do Revez. Liming increased soil pH and exchangeable Mg, in Herdade da Criméia. The SS did not pollute the soils with heavy metals. The bacterial populations of faecal origin increased immediately after the SS application, but progressively decreased as time went by, reaching values similar to the controls.

Em vastas áreas do Alentejo, os solos sob pastagem natural apresentam baixa fertilidade. A aplicação de lama residual urbana (LRU) veicula matéria orgânica (M.O.) e nutrientes para o solo, mas também pode introduzir metais pesados e bactérias de origem fecal, pelo que é conveniente monitorizar o solo após a adição destes resíduos. Comparam-se as fertilizações orgânica com LRU e a mineral, nos efeitos em alguns índices de fertilidade e contaminação metálica e fecal de um solo derivado de xistos e grauvaques, no Alentejo, no 1º ano de um campo experimental com pastagens. O campo, com um esquema experimentalem “split-plot”, foi constituído por seis talhões de 0,5 ha, correspondentes a três tratamentos de fertilização (nula, mineral e orgânica, com LRU), em dois tipos de pastagem, natural e semeada. Aplicaram-se cerca de 13 t/ha de uma mistura de LRU das ETARs de tratamento secundário de Alvito e de Vila Nova de Baronia, com teores apreciáveis de M.O., N e Ca. A adubação incluiu N, P, K, Zn e Mo. Determinaram-se os valores de pH (H2O) e os teores de M.O. total, N total, P e K “assimiláveis”, catiões de troca e de Cd, Cr, Cu, Ni, Pb e Zn extraíveis por água régia, em amostras de terra (fracção < 2 mm), colhidas à profundidade de 0-15/20 cm, antes da aplicação dos fertilizantes e cerca de um ano após a incorporação dos mesmos no solo. Para a prospecção de indicadores microbianos de contaminação fecal (bactérias coliformes e Enterococcus), efectuaram-se três amostragens de terra (0-5/10 cm), imediatamente após a aplicação de LRU e seis e 13 meses depois, nas modalidades com e sem aplicação do resíduo. Para ambos os tipos de pastagem, a aplicação de LRU beneficiou nitidamente a camada superficial do solo quanto ao teor de M.O., um ano após a aplicação dos fertilizantes, não tendo alterado o nível inicial de metais pesados ou de bactérias de origem fecal.

In wide areas of Alentejo, soils under natural pasture have low fertility. Urban biosolids (UB) application introduce organic matter (O.M.) and nutrients in the soil, but it can also add heavy metals and bacteria of faecal origin. Thus, soil monitoring after the application of these residues is required. Organic with UB and mineral fertilisations were compared regarding their effects on some fertility and metallic and faecal pollution indicators of a soil derived from schists and grauwacks, in the 1st year of a field experiment with pastures. The experimental layout was a split-plot design, with six plots of 0.5 ha, referring to three fertilisation treatments (“nil”, mineral, and organic, with UB), in natural and sown pastures. The UB application was about 13 t/ha of a UB mixture from the Alvito and Vila Nova de Baronia plants, rich in O.M., N, and Ca. Mineral fertilisation included N, P, K, Zn, and Mo. Soil samples (fraction <2 mm) collected at the 0-15/20 cm depth, before the fertilisers application and around one year after the establishment of the experiment were analysed for pH (H2O) value and O.M., total N, available P and K, exchangeable cations, and extractable aqua regia Cd, Cr, Cu, Ni, Pb, and Zn. Microbial indicators of faecal contamination (coliform bacteria and enterococci) were also evaluated from topsoil samples (0-5/10 cm), collected immediately after the UB application, and 6 and 13 months afterwards, in the treatments with and without UB application. For both types of pasture, the UB application significantly improved the O.M. content in the soil surface layer, one year after the fertiliser application, and did not change the initial soil levels of heavy metals and bacteria populations of faecal origin.

Resumo A situação do pescado no Estado do Amazonas é analisada, sendo relacionados dados referentes à produção, comercialização, industrialização e consumo deste produto, principalmente na cidade de Manaus. É apresentada uma lista dos peixes de produção significativa e a variação que ocorre durante as épocas de cheia e vazante dos rios. Ainda são discutidos: métodos empregados na pesca, condições e manuseio do pescado desde a sua captura até o mercado consumidor.

Summary This is an analysis of the situation of fishing in the State of Amazonas, giving information about the production, commercialization, and the consumption of fish, principally for the city of Manaus. A list is given of the fish most frequently found in the markets and the variation in the supply which occurs between the seasons with high water level and low water level. Methods used in fishing, and conditions, and handling of fish from the time of capture until they reach the consumer market are also discussed.