Optimizing mass production of Trichoderma asperelloides by submerged liquid fermentation and its antagonism against Sclerotinia sclerotiorum.

Commercial products based on Trichoderma are obtained mainly from solid-state fermentation. Submerged liquid fermentation is the most appropriate method compared to the solid medium for large-scale production of Trichoderma spp. The present study aimed to optimize the combination of key variables that influence the liquid fermentation process of Trichoderma asperelloides LQC-96 for conidial production coupled with its efficiency in the control of Sclerotinia sclerotiorum. In addition, we verified whether the optimized culture conditions can be used for the conidial production of Trichoderma erinaceum T-12 and T-18 and Trichoderma harzianum T-15. Fermentation studies were performed in shake flasks following a planned experimental design to reduce the number of tests and consumable costs. The effect of temperature, pH, photoperiod, carbon:nitrogen ratio and water activity on conidial production were assessed, which of pH was the only meaningful factor contributing to increased conidial production of T. asperelloides LQC-96. From the five variables studied initially, pH and C:N ratio were further used in the second design (rotational central composite design-RCCD). Hence, the best conditions for the production of T. asperelloides LQC-96 conidia by liquid fermentation consisted of initial pH of 3.5, C:N ratio of 200:1 at 30 °C, without glycerol, and under 24 h photoperiod. The highest conidial concentration was observed after seven days of fermentation. Under these optimal conditions, T. erinaceum T-12 and T-18, and T. harzianum T-15 were also cultivated, but only LQC-96 efficiently parasitized S. sclerotiorum, precluding sclerotium myceliogenic germination. Our findings propose optimal fermentation conditions that maximize conidial production of T. asperelloides as a potential biofungicide against S. sclerotiorum.

The impact of an aquatic exercise program on BDNF levels in Parkinson's disease patients: short-and long-term outcomes.

The present study aimed to analyze the short-and long-term effects of an aquatic exercise program on plasma brain-derived neurotrophic factor (BDNF) levels in individuals with Parkinson's disease (PD). The aquatic exercise program lasted one month, and consisted of two sessions per week (1 hour per session). Blood samples were collected at four different timepoints: pre-intervention (T0), immediately after the first session (T1), 48 hours after the first session (T2), and 1 month after the intervention (T3). We found a significant decrease in BDNF levels at T2 vs T1 (p<0.05). However, no changes were observed at the other time-points. Our results demonstrated that the intervention reduced plasma BDNF levels in PD individuals in a time-dependent manner: specifically, we observed acute effects, but no delayed effects.

The biochemical characterization of two phosphate transport systems in Phytomonas serpens.

Inorganic phosphate (Pi) is an essential nutrient for all organisms because it is required for a variety of biochemical processes, such as signal transduction and the synthesis of phosphate-containing biomolecules. Assays of 32Pi uptake performed in the absence or in the presence of Na+ indicated the existence of a Na+-dependent and a Na+-independent Pi transporter in Phytomonas serpens. Phylogenetic analysis of two hypothetical protein sequences of Phytomonas (EM1) showed similarities to the high-affinity Pi transporters of Saccharomyces cerevisiae: Pho84, a Na+-independent Pi transporter, and Pho89, a Na+-dependent Pi transporter. Plasma membrane depolarization by FCCP, an H+ ionophore, strongly decreased Pi uptake via both Na+-independent and Na+-dependent carriers, indicating that a membrane potential is essential for Pi influx. In addition, the furosemide-sensitive Na+-pump activity in the cells grown in low Pi conditions was found to be higher than the activity detected in the plasma membrane of cells cultivated at high Pi concentration, suggesting that the up-regulation of the Na+-ATPase pump could be related to the increase of Pi uptake by the Pho89p Na+:Pi symporter. Here we characterize for the first time two inorganic phosphate transporters powered by Na+ and H+ gradients and activated by low Pi availability in the phytopathogen P. serpens.

Elaboração e validação de escalas diagramáticas para avaliação da severidade da mancha-bacteriana do feijão - caupi em cultivares com trifólios morfologicamente distintos / Elaboration and validation of a diagrammatic scale to assess bacterial blight severity on cowpea cultivars with morphologically distinct leaflets

Duas escalas diagramáticas foram desenvolvidas para quantificar a severidade da mancha-bacteriana do feijão-caupi, causada pela bactéria Xanthomonas axonopodis pv. vignicola, em cultivares de feijão-caupi de trifólios com folíolos ovais e lanceolados. Trifólios com diferentes níveis de severidade foram coletados para determinação do limite mínimo e máximo de severidade, sendo níveis intermediários determinados de acordo com a "Lei do Estímulo de Weber-Fechner". Os níveis propostos foram: 0,05; 0,25; 1,00; 3,40; 9,70; e 17,70% para trifólios com folíolos ovais e 0,10; 0,35; 1,10; 3,50; 9,30; e 19,10% para trifólios com folíolos lanceolados. As escalas foram validadas por meio do cálculo da sua acurácia e precisão. Para tanto, realizou-se análises de regressão linear da severidade real com a estimada com e sem o uso das escalas propostas. As estimativas foram feitas por 12 avaliadores sem experiência em quantificar essa doença em 50 trifólios com diferentes níveis de severidade da mancha-bacteriana. Para isso, avaliou-se a acurácia e a precisão dos avaliadores. Com a utilização das escalas diagramáticas, obtiveram-se níveis satisfatórios de acurácia e precisão. Assim, as escalas propostas tornam-se uma ferramenta fácil e de rápida utilização, que podem ser adotadas para auxiliar na avaliação da severidade da mancha-bacteriana do feijão-caupi, em estudos epidemiológicos e seleção de cultivares resistentes a doença.

Two diagrammatic scales were developed to assess the severity of bacterial blight of cowpea, caused by Xanthomonas axonopodis pv. vignicola in cowpea cultivars with oval and lanceolate leaflets, considering the limits for minimum and maximum severity of the disease, as well as intermediate levels. The levels were determined in accordance with the "Weber-Fechner's Stimulus Responses Law", representing: 0.05; 0.25; 1.00; 3.40; 9.70; and 17.70% for diagrammatic scale with oval leaflets and 0.10; 0.35; 1.10; 3, 50; 9.30; and 19.10% for lanceolate leaflets. The scales were validated by means of their accuracy and precision. To calculate these factors, linear regression analysis of actual disease severity and the estimated ones with and without the use of the scales. Twelve non-experienced evaluators estimated 50 leaflets with different levels of severity bacterial blight. For this, the accuracy and precision of the evaluators was assessed. With the use of diagrammatic scales, there were satisfactory levels of accuracy and precision. Thus, the diagrammatic scales proposed were considered appropriate to assess the severity of bacterial blight of cowpea and can be used in epidemiological studies, to evaluate the reactions of genotypes to the disease.

Leishmania amazonensis: inhibition of 3'-nucleotidase activity by Cu2+ ions.

Free Cu(2+) is toxic due to the capacity of free copper ions to catalyze the production of reactive oxygen species (ROS) that can modify the structure and/or function of biomolecules. In addition, non-specific binding to enzymes, which modifies their catalytic activities, can occur. In this work, the mechanisms underlying the ability of copper to inhibit 3'-nucleotidase from Leishmania amazonensis (La3'-nucleotidase) were investigated. To that end, La3'-nucleotidase activity was assayed with CuCl(2) in the presence of ascorbate or hydrogen peroxide to discriminate non-specific binding effects from pro-oxidant effects of copper. Copper inhibitory effects were greater at more acidic pH than at alkaline pH. The addition of enzyme substrate, adenosine 3'-monophosphate (3'AMP), prevented the inhibition of enzyme activity by copper. Thiol-containing compounds were able to protect the enzyme activity against inhibition due to copper. The specific copper chelating agent bathocuproine sulphonate (BCS) restored enzyme activity after pre-treatment of the enzyme with copper. La3'-nucleotidase activity was found to be resistant to ROS generated during oxidation reactions of ascorbate and hydrogen peroxide catalyzed by copper. Our results suggest that Cu(2+) ions exert their inhibitory effects by binding to specific motifs of the 3'-nucleotidase protein and that the enzyme appears to be extremely resistant to ROS.

Leishmania amazonensis: characterization of an ecto-3'-nucleotidase activity and its possible role in virulence.

Ecto-3'-nucleotidase/nuclease (3'NT/NU) is a membrane-bound enzyme that plays a key role in the nutrition of Leishmania sp. protozoan parasites. This enzyme generates nucleosides via hydrolyzes of 3'mononucleotides and nucleic acids, which enter the cell by specific transporters. In this work, we identify and characterize Leishmania amazonensis ecto-3'-nucleotidase activity (La3'-nucleotidase), report ammonium tetrathiomolybdate (TTM) as a novel La3'-nucleotidase inhibitor and approach the possible involvement of ecto-3'-nucleotidase in cellular adhesion. La3'-nucleotidase presented characteristics similar to those reported for the class I single-strand nuclease family; a molecular weight of approximately 40 kDa and optimum activity in an alkaline pH range were observed. Although it is conserved among the genus, La3'-nucleotidase displays different kinetic properties; it can be inhibited by vanadate, molybdate and Cu(2+) ions. Interestingly, ecto-3'-nucleotidase activity is 60-fold higher than that of ecto-5'-nucleotidase in L. amazonensis. Additionally, ecto-3'-nucleotidase activity is two-fold higher in virulent L. amazonensis cells than in avirulent ones. Notably, macrophage-parasite attachment/invasion was increased by 400% in the presence of adenosine 3'-monophosphate (3'AMP); however, this effect was reverted by TTM treatment. We believe that La3'-nucleotidase may play a significant role in the generation of adenosine, which may contribute to mammalian host immune response impairment and establishment of infection.

Ocorrência de Passalora bougainvilleae (Muntañola) Castañeda & Braun associado à Bougainvillea spectabilis Willd. em Boa Vista, Roraima / Repot of Passalora bougainvilleae (Muntañola) Castañeda & Braun associated with Bougainvillea spectablis Willd. in Boa Vista, Roraima

O fungo Passalora bougainvilleae é relatado, pela primeira vez, na região de Boa Vista, Roraima, associado a manchas foliares na planta ornamental Bougainvillea spectabilis. As características do fungo são conidióforos agregados em fascículos, emergindo de um estroma subcuticular na face abaxial de lesões velhas. Os conidióforos são lisos, retos, de coloração marrom, não ramificados, maioria asseptados, medindo de 26-57µm x 4µm. Células conidiogênicas terminais de proliferação simpodial com cicatrizes escuras e pouco espessas. Os conídios são solitários, marrom claro, obclavados, retos a ligeiramente curvos, medindo de 32-70 µm x 4-5 µm, maioria com 3 septos, apresentando um hilo truncado e ápice obtuso.

The fungi Passalora bougainvilleae is reported, for the first time, Boa Vista, Roraima state, causing leaf spots on ornamental specie Bougainvillea spectabilis. The fungi characteristics are conidiophores hypophyllous in fascicles arising from the upper cells of a stroma subcuticular at old spots. The conidiphores are smooth, straight, pale brown, non-branched, mostly aseptate, 26-57µm x 4µm. Conidiogenous cells terminal, sympodial with black and thin scars. Conidia solitary, pale brown, obclavate, straight to slightly flexuous, 32-70 µm x 4-5 µm, 3 septate, rounded at apex, truncate at base, with a conspicuous hilum.

Podridão-mole em plantas de cebolinha causada por Pectobacterium carotovorumsubsp. carotovorum em Roraima / Soft rot of bunching onion plants caused by Pectobacterium carotovorumsubsp. carotovorum in Roraima, Brazil

A ocorrência de Pectobacterium carotovorum subsp. carotovorum (=Erwinia carotovora subsp. carotovora) em cebolinha (Allium fistulosum) é relatada pela primeira vez na região norte do Brasil. Até então sua ocorrência estava registrada apenas no Distrito Federal.

This is the first report of Pectobacterium carotovorum subsp. carotovorum (=Erwinia carotovora subsp. carotovora) causing soft rot of bunching onion (Allium fistulosum) plants in Roraima, Brazil. Its occurrence is reported only in Distrito Federal.