Differential activity of a lectin from Solieria filiformis against human pathogenic bacteria

A lectin isolated from the red alga Solieria filiformis was evaluated for its effect on the growth of 8 gram-negative and 3 gram-positive bacteria cultivated in liquid medium (three independent experiments/bacterium). The lectin (500 µg/mL) stimulated the growth of the gram-positive species Bacillus cereus and inhibited the growth of the gram-negative species Serratia marcescens, Salmonella typhi, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus sp, and Pseudomonas aeruginosa at 1000 µg/mL but the lectin (10-1000 µg/mL) had no effect on the growth of the gram-positive bacteria Staphylococcus aureus and B. subtilis, or on the gram-negative bacteria Escherichia coli and Salmonella typhimurium. The purified lectin significantly reduced the cell density of gram-negative bacteria, although no changes in growth phases (log, exponential and of decline) were observed. It is possible that the interaction of S. filiformis lectin with the cell surface receptors of gram-negative bacteria promotes alterations in the flow of nutrients, which would explain the bacteriostatic effect. Growth stimulation of the gram-positive bacterium B. cereus was more marked in the presence of the lectin at a concentration of 1000 µg/mL. The stimulation of the growth of B. cereus was not observed when the lectin was previously incubated with mannan (125 µg/mL), its hapten. Thus, we suggest the involvement of the binding site of the lectin in this effect. The present study reports the first data on the inhibition and stimulation of pathogenic bacterial cells by marine alga lectins.

Antinociceptive properties in mice of a lectin isolated from the marine alga Amansia multifida Lamouroux

The antinociceptive effects of a lectin (LEC) isolated from the marine alga Amansia multifida were determined in Swiss mice. The LEC (1, 5, and 10 mg/kg) inhibited acetic acid-induced abdominal writhings in a dose-dependent manner after intraperitoneal or oral administration. A partial but significant inhibition of writhings was observed after the combination of LEC (10 mg/kg) with avidin (1 mg/kg), a potent inhibitor of the hemmaglutinant activity of the lectin. However, total writhing inhibition was demonstrable in the group of mice treated with LEC plus mannose (1 mg/kg), as compared to LEC alone or to control groups. Furthermore, avidin and mainly mannose also play a role in antinociception, somehow facilitating the interaction of LEC with its active cell sites. In the formalin test, although both phases of the response were significantly inhibited, the effect of LEC was predominant during phase 2, causing inhibition of licking time that ranged from 48 to 88% after oral (5 and 10 mg/kg) and intraperitoneal (1 to 5 mg/kg) administration. As is the case with morphine, the effect of LEC (2 mg/kg) was reversed by naloxone (2 mg/kg), indicating the involvement of the opioid system. LEC was also effective in the hot-plate test, producing inhibitory responses to the thermal stimulus, and its effects were blocked by naloxone. In the pentobarbital-induced sleeping time, although LEC did not alter the onset of sleep significantly, it increased the time of sleep within the same dose range compared to control. These results show that LEC presents antinociceptive effects of both central and peripheral origin, possibly involving the participation of the opioid system.

High levels of serum mannose-binding lectin are associated with the severity of clinical signs of leptospirosis

The clinical heterogeneity observed in leptospirosis may be associated with host factors or bacteria virulence. Human serum mannose-binding lectin (MBL) recognizes many pathogens, and low levels of this lectin are associated with susceptibility to infection. MBL is also implicated in the modulation of the inflammatory process. We determined the levels of serum MBL during leptospirosis infection. A double-antibody sandwich ELISA was used to detect the immunoreactive serum MBL. The ELISA plates were coated with monoclonal antibody to MBL and bound MBL or recombinant human MBL were detected by rabbit anti-human MBL serum. HRPO-conjugated goat anti-rabbit antibody was used for detection of the reaction. Two groups of patients seen at referral hospitals in Recife, PE, Brazil, were divided according to the year of infection, 2001 (N = 61) or 2002 (N = 57) and compared in terms of disease severity and levels of serum MBL. A group of healthy volunteers (N = 97) matched by age, gender, and ethnic background was used as control. Patients infected in 2001 had more severe outcomes than those infected in 2002, including jaundice, hemorrhage, respiratory alteration, and renal complication (P = 0.0009; chi-square test). The frequency of patients producing serum MBL >1000 ng/mL was higher in the 2001 group than in the 2002 and control groups (P < 0.01), suggesting an association of MBL level with disease severity. The involvement of MBL and genetic variation of the MBL2 gene should be further evaluated to establish the role of this lectin in the pathogenesis of leptospirosis.

Purification, partial characterization and antimicrobial activity of Lectin from Chenopodium Quinoa seeds

Abstract A novel lectin was isolated from the seeds of Chenopodium quinoa. To achieve this end, the crude extract from the quinoa was submitted to two purification steps, Sephadex G50 and Mono Q. The hemagglutinating activity showed that this lectin agglutinates human erythrocytes. Its activity is inhibited by glucose and mannose, and remained stable under a wide range of pH levels and temperatures. The quinoa lectin was found to be a heterodimeric lectin of approximately 60 kDa, consisting of two subunits of approximately 25 kDa and 35 kDa. This lectin had its antimicrobial activity tested against several bacteria strains and effectively inhibited three strains. These strains were all Gram-negative, making this lectin a promising antimicrobial tool.

Production of cabbage grown in pots containing legumes' root and shoot

Roots effect is not generally considered in studies assessing the performance of crops in response to green manuring. However, such effect can contribute to a better understanding of crop rotation. The aim of this study was to assess the effect of root and shoot of two legumes on the production of cabbage. The experiment was conducted in pots of 10 liters containing substrate of 2:1 soil/sand. The experiment was arranged in a factorial scheme (2x3 + 2) in a randomized block design with five replicates using two legume species (Crotalaria juncea L. and Canavalia ensiformis L), three plant parts (root, shoot, or whole plant), and two additional treatments (mineral fertilization with 100% and 50% of the recommended dose of N for growing cabbage). Pots with legume treatments received mineral fertilizer with 50% of the recommended dose of N for growing cabbage. The experimental plot consisted of a pot containing one plant of cabbage. Legumes were grown in pots and harvested at 78 days. The root biomass was determined in extra pots. Production was assessed using head fresh and dry weight. The application of the whole plant of both legume species reduced cabbage production. However, root or shoot of both legume species was equivalent to 50% of mineral N fertilization required for the cultivation of cabbage.

Susceptibilidade de espécies de plantas com potencial de fitorremediação do herbicida sulfentrazone

Dentre as novas tecnologias, a fitorremediação é opção para a descontaminação de áreas que receberam intensas aplicações de herbicidas. Este trabalho foi desenvolvido com o objetivo de selecionar espécies com potencial para a fitorremediação de solos contaminados com o herbicida sulfentrazone. As espécies testadas foram: Calopogonium mucunoides, híbrido de sorgo (Sorghum bicolor x Sorghum sudanense), Crotalaria breviflora, Crotalaria juncea, Canavalia ensiformis, Dolichos lablab, Stizolobium deeringianum e Stizolobium aterrimum, cultivadas em cinco doses do sulfentrazone (0, 200, 400, 800 e 1.600 g ha-1). O experimento foi instalado em casa de vegetação, utilizando-se vasos com capacidade para 6 L, preenchidos com amostras de solo coletadas na profundidade de 0-20 cm. Foram avaliadas a fitotoxicidade do herbicida, a altura de plantas e a massa de matéria seca da parte aérea e de raízes. As espécies que manifestaram os menores sintomas de fitotoxicidade e os maiores índices de altura e matéria fresca e seca foram C. juncea, C. ensiformis e D. lablab, sendo selecionadas como tolerantes ao herbicida sulfentrazone e com potencial para fitorremediação.

Phytomass production and nutrient accumulation by green manure species

Green manuring is recognized as a viable alternative to improve nutrient cycling in soils. The aim of this study was to evaluate the phytomass production and nutrient accumulation in shoots of the summer green manures jack bean [Canavalia ensiformis (L.) DC.], dwarf pigeon pea (Cajanus cajanvar var. Flavus DC.), dwarf mucuna [Mucuna deeringiana (Bort) Merr] and sunn hemp (Crotalaria juncea L.), under nitrogen fertilization and/or inoculation with N-fixing bacteria. A split plot design was arranged with the four Fabaceae species as main plots and nitrogen fertilization (with and without) and inoculation with diazotrophic bacteria (with and without) as the subplots, in a 2² factorial. The experiment was arranged as a randomized complete block design with four replications. In the conditions of this trial, the sunn hemp had the highest production of shoot phytomass (12.4 Mg ha-1) and nutrient accumulation, while the dwarf mucuna had the lowest production of shoot phytomass (3.9 Mg ha-1) and nutrient accumulation. The results showed no effect of nitrogen fertilization or inoculation with N-fixing bacteria on the production of shoot phytomass and nutrient accumulation, except for inoculation without nitrogen fertilization, resulting in greater P accumulation (p <0.05) in the sunn hemp and greater Zn and Mn accumulation in the dwarf mucuna. These findings indicate that N fertilization or inoculation with N2-fixing bacteria for Fabaceae are low efficiency practices in the edaphoclimatic conditions of this study.

Weed suppression by green manure in an agroecological system

ABSTRACT Green manure promotes efficient suppression of weeds, but green manure species can exhibit distinct behaviors, depending on the environmental conditions. This study aimed to evaluate the potential of soil mulching and weed suppression by spring/summer green manure species grown in the spring/summer season, at different growth stages and after management (cut), for 90 days during the cassava crop cycle. The study was carried out in the 2010/2011 season, in a system managed under agroecological principles. The treatments consisted of different green manure species and arrangements: Crotalaria juncea, Cajanus cajan, Canavalia brasiliensis, Canavalia ensiformis, Pennisetum americanum, Crotalaria juncea and Pennisetum americanum intercropped; Mucuna aterrima, Sorghum bicolor, a mixture of all the green manures in study and a control plot under fallow. The experiment was arranged in a randomized block design with four replications. The evaluations of the soil cover either by the green manures or weeds were performed at 45, 90 and 105 days after the emergence of the green manures. The cassava crop was planted under reduced tillage system at 11 days after the cut of the green manures. The percentage of soil covered by weeds and the dry matter produced were evaluated at 30, 60 and 90 days after planting. The results showed that the green manures had a suppressive effect on weeds during their life cycle, as well as during the first months after its management (cut), composing the mulch.

Lectins for the detection of IgM antibodies to T. gondii in the diagnosis of acute toxoplasmosis by immunofluorescence test

Lectins were labeled with fluorescein and tried as conjugates in the immunofluorescence (IP) test for the detection of IgM antibodies to T. gondii, in the diagnosis of acute toxoplasmosis. This approach was an attempt to find alternative reagents for anti-human IgM fluorescent conjugates (AHIgMFC), which contain quite frequently anaibcdies to toxoplasma, as contaminants, due to natural T. gondii infections among animals used for imunization. Lentil (Lens culinaris) lectin fluorescence conjugates (LcFC) provided most satisfactory results. The evaluation of LcFC carried out in a total of 179 sera from patients with acute and chronic toxoplasmosis, with non-related infections or healthy subjects, gave high values of relative efficiency, co-positivity and co-negativity indices, respectively 0.989, 0.969 and 1.000, in reference to the conventional AHIgMFC. Moreover, three batches of LcFC successively prepared gave reproducible test results. The advantage of LcFC as an alternative reagent for the serodiagnosis of acute toxoplasmosis is supported by practical aspects of its preparation.

The diagnostic importance of species specific and cross-reactive components of Taenia solium, Echinococcus granulosus, and Hymenolepis nana

Sera from patients infected with Taenia solium, Hymenolepis nana and Echinococcus granulosus were tested against homologous and heterologous parasite antigens using an ELISA assay, and a high degree of cross-reactivity was verified. To identify polypeptides responsible for this cross reactivity, the Enzyme Linked Immunoelectro Transfer Blot (EITB) was used. Sera from infected patients with T.solium, H.nana, and E.granulosus were assessed against crude, ammonium sulphate precipitated (TSASP), and lentil-lectin purified antigens of T.solium and crude antigens of.H.nana and E.granulosus. Several bands, recognized by sera from patients with T.solium, H.nana, and E.granulosus infections, were common to either two or all three cestodes. Unique reactive bands in H.nana were noted at 49 and 66 K-Da and in E.granulosus at 17-21 K-Da and at 27-32 K-Da. In the crude cysticercosis extract, a specific non glycoprotein band was present at 61-67 K-Da in addiction to specific glycoprotein bands of 50, 42, 24, 21, 18, 14, and 13 K-Da. None of the sera from patients with H.nana or E.granulosus infection cross reacted with these seven glycoprotein bands considered specific for T.solium infection.

The influence of carbohydrates in the interaction of Paracoccidioides brasiliensis with CCL-6 cells in vitro

INTRODUCTION: Little is known about the early events in the interaction between Paracoccidioides brasiliensis and its host. To understand the effect of carbohydrates in the interaction between the fungus and epithelial cell in culture, we analyzed the influence of different carbohydrate solutions on the adhesion of P. brasiliensis yeast cells to CCL-6 cells in culture. METHODS: Fungal cells were cultivated with the epithelial cell line, and different concentrations of D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine, D-galactosamine, sorbitol and fructose were added at the beginning of the experiment. Six hours after the treatment, the cells were fixed and observed by light microscopy. The number of P. brasiliensis cells that were adhered to the CCL-6 monolayer was estimated. RESULTS: The number of adhesion events was diminished following treatments with D-fucose, N-acetyl-glucosamine, D-mannose, D-glucosamine and D-galactosamine as compared to the untreated controls. Sorbitol and fructose-treated cells had the same adhesion behavior as the observed in the control. P. brasiliensis propagules were treated with fluorescent lectins. The FITC-labeled lectins WGA and Con-A bound to P. brasiliensis yeast cells, while SBA and PNA did not. CONCLUSIONS: The perceptual of adhesion between P. brasiliensis and CCL-6 cells decreased with the use of D-mannose, N-acetyl-glucosamine and D-glucosamine. The assay using FITC-labeled lectins suggests the presence of N-acetyl-glucosamine, α-mannose and α-glucose on the P. brasiliensis cell surface. An enhanced knowledge of the mediators of adhesion on P. brasiliensis could be useful in the future for the development of more efficient and less harmful methods for disease treatment and control.

Avaliação de espécies leguminosas na formação de cafezais no segmento da agricultura familiar no Acre

Leguminosas quando consorciadas com o café e usadas como adubação verde podem contribuir fornecendo nitrogênio e proteção ao solo pela adição de matéria orgânica. O trabalho foi conduzido com o objetivo de avaliar o efeito do uso de leguminosas no sistema de produção de café, no segmento de agricultura familiar, visando promover a implantação e a manutenção dos cafezais de forma técnica e economicamente sustentável. O experimento foi conduzido no período de novembro de 2000 a abril de 2003, em delineamento experimental de blocos ao acaso em esquema de parcelas subdivididas, com quatro repetições. Os tratamentos utilizados nas parcelas foram as leguminosas (Stizolobium atterrimum, Cajanus cajan, Canavalia endiformis e Flemingia congesta) e a testemunha (sem leguminosa) e, nas subparcelas, duas doses de N (0 e 22 g de N por cova). A Flemingia congesta e a Mucuna aterrima foram as leguminosas que mais influenciaram positivamente a produtividade dos cafeeiros, independente da adubação nitrogenada. Em relação à testemunha, o aumento em produção foi de 109% quando utilizou-se a Flemingia congesta e 52% com a Mucuna aterrima. A Flemingia congesta foi também a leguminosa que melhor controlou as invasoras, dado o volume de fitomassa produzida e a possibilidade de 2 cortes durante um período de doze meses, evidenciando o potencial desta leguminosa na formação de novos cafezais no Acre. Por outro lado, a Canavalia ensiformis, leguminosa que é normalmente utilizada nas entrelinhas dos cafeeiros pelos cafeicultores, do Acre e de outras regiões produtoras, neste trabalho influenciou negativamente a altura das plantas, diâmetro da copa e crescimento dos cafeeiros.

Produção de matéria seca e eficiência nutricional para P, Ca e Mg em leguminosas herbáceas

As leguminosas para adubação verde têm sido introduzidas nos sistemas agrícolas para a recuperação de solos desgastados pelo uso intensivo. O objetivo desse trabalho foi avaliar a resposta de leguminosas herbáceas à aplicação de calcário e fósforo sobre a produção de biomassa e eficiência nutricional para o P, Ca e Mg. O experimento foi desenvolvido em casa-de-vegetação do Departamento de Ciência do Solo da Universidade Federal Rural da Amazônia, utilizando-se um Latossolo Amarelo distrófico coletado na profundidade de 0 - 20 cm, em Belém, PA. O delineamento experimental usado foi o de blocos ao acaso, com arranjo de tratamentos em um fatorial 3³, onde foram comparadas três espécies de leguminosas: mucuna preta (Stizolobium atterrimum), mucuna cochinchinensis (Stilozobium cochinchinensis) e feijão-de-porco (Canavalia ensiformes); três níveis de calagem: de calcário dolomítico (0, 4 e 8 t.ha-1) e de fósforo (0, 25 e 45 mg.dm-3), com quatro repetições cada um. Mucuna cochinchinensis e o feijão de porco produziram maiores quantidades de biomassa vegetal do que a mucuna preta, sendo que a primeira espécie acumulou maior quantidade de P e a segunda maior quantidade de Ca. Mucuna cochinchinensis apresentou maior eficiência de utilização de P, Ca e Mg na ausência da calagem. Nos solos intemperizados com baixa concentração de P, Ca e Mg, o feijão-de-porco e a mucuna cochinchinensis poderão ter melhor desempenho do que a mucuna preta, visto que apresentaram maior eficiência de translocação e de utilização desses nutrientes, respectivamente.

Avaliação de indicadores de qualidade do solo sob alternativas de recuperação do solo no Nordeste Paraense

Os indicadores convencionais de fertilidade de solo, mais comumente utilizados, enfatizam o balanço ácido-base e o conteúdo de elementos químicos, mas não consideram adequadamente as propriedades biológicas do solo, responsáveis por grande parte do seu funcionamento. Os indicadores biológicos, ainda pouco difundidos, levam em consideração a atividade microbiana do solo que é mais sensível à dinâmica do solo. Em Marapanim-PA, e utilizando um desenho experimental inteiramente casualizado, com quatro repetições, avaliou-se o comportamento dos indicadores do solo carbono e nitrogênio da biomassa microbiana, fosfatase ácida, matéria orgânica, carbono orgânico e fósforo disponível, em um solo com cultivo sequencial de mandioca e milho, preparo com corte-e-trituração da capoeira original. Os fatores corte-e-trituração, fosfato natural e adubação verde foram arranjados nos seguintes tratamentos: capoeira; testemunha; fosfato natural (FN); feijão-de-porco [Canavalia ensiformis (L.) DC] (FP); FN + FP; FN + guandu [Cajanus cajan (L.) Millspaugh]. (G); FN + titônia [Tithonia diversifolia (Hemsley) A. Gray] (T); FP + G; FP + T. Em quatro épocas de amostragens, foram tiradas amostras compostas do solo (0 - 10 cm) de cada parcela para análise dos indicadores. Os indicadores estudados foram influenciados pela época de amostragem do solo. Os indicadores fosfatase ácida, nitrogênio da biomassa microbiana e carbono da biomassa microbiana foram capazes de detectar os efeitos de maior número de tratamentos de recuperação do solo testados, enquanto a matéria orgânica do solo foi o menos capaz. Ao longo do tempo, a resposta dos indicadores variou em função dos fatores estudados.

Grasses and legumes as cover crop in no-tillage system in northeastern Pará Brazil

Studies to select one or more species of coverage plants adapted to Amazonian soil and climate conditions of the Amazon are a promising strategy for the improvement of environmental quality, establishing no-till agricultural systems, and thereby reducing the impacts of monoculture farming. The aim of this study was to assess the persistence time, half-life time, macronutrient content and accumulation, and C:N ratio of straw coverage in a Ultisol in northeastern Pará. Experimental design was randomized blocks with five treatments and five replicates. Plants were harvested after 105 days, growth and biomass production was quantified. After 84 days, soil coverage was 97, 85, 52, 50, and 15% for signalgrass (Brachiaria brizantha) (syn. Urochloa), dense crowngrass (Panicum purpurascens), jack bean (Canavalia ensiformes), pearl millet (Pennisetum americanum) and sunn hemp (Crotalaria juncea,), respectively. Signalgrass yielded the greatest dry matter production (9,696 kg ha-1). It also had high C:N ratio (38.4), long half-life (86.5 days) and a high persistence in the field. Jack bean also showed high dry matter production (8,950 kg ha-1), but it had low C:N ratio (17.4) and lower half-life time (39 days) than the grasses. These attributes indicate that signalgrass and jack bean have a high potential for use as cover plants in no-till agricultural systems in the State of Pará.