Quitosana na indução de resistência ao tombamento de plântulas de espécies olerícolas e no controle de fitopatógenos in vitro

Induction of resistance is defined as the activation of a state of resistance against diseases which is induced systemically in plants by the use of biotic or abiotic agents without any modification of the plant genome, occurring non-specific way, by activating genes coding for various plant defense responses. Chitosan is a polymer derived from the deacetylation of chitin, which is found in large quantities in crustacean shell, and studied with the potential to control plant pathogens, both by its direct fungistatic action, as the ability to induce protection of plants, indicating the presence of molecules of elicitoras characteristics. Three experiments with objective of evaluating the potential of chitosan in the seedling resistance induction were developed, beet (Beta vulgaris) seeds, cucumber (Cucumis sativus) seeds and tomato (Solanum lycopersicum) seeds, and the control of Fusarium sp., Rhizoctonia solani K¨uhn e Pythium sp. in vitro conditions. The experimental design was completely randomized, with four replications. Beet seeds, tomato and cucumber were submerged in chitosan solution for 20 minutes, in concentrations of 0.25, 0.5, 1 and 2% in the control and distilled water. Seeds were sown in trays containing Plantmax Florestalr substrate sterilized and inoculated with Fusarium sp., Rhizoctonia solani K¨unh and Pythium sp., respectively for the three cultures. The experiment was conducted for 14 days in growth chamber with controlled temperature (25 C 2 C), light (12 hour photoperiod) and humidity (70% 10%). The evaluations were seed emergency, seedling damping-off, seedling length, fresh weight and activity of the enzymes phenylalanine amˆonia-liase (PAL), chitinase and b-1,3-glucanase. It was also rated the mycelial growth of Fusarium sp., Pythium sp. and R. solani on P.D.A. (Potato-Dextrose and Agar) culture medium containing chitosan at the same concentrations evaluated in seeds. For beet growing, seed treatment with chitosan presented higher emergence and the length of the seedlings, and reduced the percentage of tipping. Treatment with chitosan activated the systemic acquired resistance with expression of chitinase and b-1,3-glucanase enzymes. For the tomato crop in chitosan concentration of 0.25% favored the emergency of seedlings, reduced the incidence of tipping and activated the PAL enzymes, chitinase and b-1,3-glucanase. In cucumber on the concentration of up 0.5% favored seedlings emergence and reduces the incidence of tipping. Chitosan activated the PAL enzymes and b-1,3-glucanase. Chitosan also presented fungistatic action on the initial growth of Pythium sp. and R. solani in vitro conditions, however, such action did not prevail until the end of the experiment. To Fusarium sp. the concentration of chitosan resulted in the reduction of mycelial growth in vitro.

Crescimento micelial de Ganoderma lucidum em diferentes substratos e relação com concentração de β-Glucanas, atividade antioxidante e efeitos sobre o desempenho de coelhos

Ganoderma lucidum is a medicinal mushroom traditionally consumed in Asian countries that presents several beneficial effects already verified. Despite all studies about their bioactive compounds, the best cultivation media enrichment aiming to increase the production of these compounds is still uncertain. Besides, few studies are related to the performance of production animals. In order to test different cultivation media for G. lucidum mycelia, agricultural residues (solid state fermentation) and different sugars and aminoacids (in culture media with agar), were tested to evaluate G. lucidum mycelium growth. Supply of flour with G. lucidum mycelia obtained by solid state fermentation (wheat grain) for rabbits was also evaluated. Mycelium of G. lucidum developed very well in all agricultural residues, soybean hulls was the residue that presented higher growth rate and higher concentration of β-glucans. In the cultivation media experiment, G. lucidum also developed well, media that contained cellobiose and tyrosine, despite presenting lower growth rates and total growth within 10 days, produced mycelia with higher concentration of β-glucans and trolox equivalent antioxidant capacity (TEAC), respectively. Rabbits did not show any sign of intolerance to feed with different concentrations of flour with G. lucidum mycelium. All performance parameters and dressing percentages were adequate to the age at which they were slaughtered. Histological evaluation of organs presented alterations in renal cells (tubular and glomerulus), indicating a possible renal lesion according to the increase of flour with mycelium in feed. Histomorphometric evaluation showed increased vilous height in ileum and decreased vilous width of jejunus at 0.5% concentration, and decrease in crypt diameter according to the increase of concentration of flour with mycelium in feed. These results indicate the possibility of more studies regarding the aspects about cellobiose and tyrosine utilization for the production of bioactive compounds, and about toxicity of this mushroom mycelia, assuring the safety in supplying this product for animails.

Potencial do óleo de Aloysia citriodora Palau no controle de fitopatógenos e na indução de resistência ao tombamento de plântulas de feijão, pepino e beterraba

The crops are affected by pests and diseases that decrease productivity. Among them are the damping off of seedlings that can occur in pre and post-emergence. In bean crops, cucumber and beet these diseases occur, being caused by various pathogens, especialy fitopathogenic fungi. Several measures are used for the controle of such diseases, among them, is the chemical seed treatment fungicides. However, society has become increasingly concerned about the quality and food and environmental contamination, generation a growting search for sensitive products to humans and the environment. The use of essential oils to control plant pathogens is an example of alternative tested by science in the search for less aggressive technologies. This study aimed to evaluate the efficiency of the use of essential oil Aloysia citriodora, in control of pathogens causing damping off in beans, cucumber and beet. This thesis was divided in four chapters, the introductory first, and the other addressing the control of Pythium sp. in beans, Sclerotinia sclerotiorum on cucumber, and Fusarium sp. on beet. The methodology consisted of four experiments in each pathosystem, with all the work done at the Federal Technological University of Parana, Campus Dois Vizinhos. In the first experiment evaluated the fungistatic and fungicidal effect of the essential oil of A. citriodora on PDA in vitro in mycelial growth of pathogens studied. In the second experiment evaluated the in vitro effect of essential oil concentrations of A. citriodora in BD medium on microscope slides, on the germination of sporangia Pythium sp. and conidia Fusarium sp., and in Petri dishes with PDA medium, the sclerotia germination speed index of S. sclerotiorum. In the third experiment, we evaluated in germination test in paper roll (PR), the phytotoxic effect or not the use of essential oil concentrations of A. citriodora in dry bean seed, cucumber and beet. The variables used to assess this experiment were the germination percentage, mediun green mass per plant and average length of seedlings. In the fourth experiment we assessed the effect of treating bean seeds, cucumber and beet with essential oil contents of A. citriodora, seeds in their subsequent substrates contamined with pathogens studied, Pythium sp., S. sclerotiorum and Fusarium sp. In this experiment we used the following variables: percentage of emergence, percentage of post-emergence damping off, green average mass per plant, average length per plant and biochemical analyzes. The biochemistry of plant tissues evaluated were as follows: protein content, enzymatic activities of peroxidases, phenylalanine ammonia-liase (PAL), chitinases and β-1,3-glucanases. The in vitro results show that the essential oil has fungistatic and fungicidal effect on mycelial growth, on sporangia germination, conidia and sclerotia of the pathogens studied in this work, wich may be related to its major components, citral and limonene. The oil also exhibits low phytotoxicity to seeds of the species studied, only in beans decreases germination in most studied dosage (0,25%), cucumber also in the higher dosage (0,25%) reduce the length of seedlings, and beet there were no negative effects to the seedlings. In the test in substrate contaminated with the pathogens, the use of essential oil: increased germination and decreased post emergence damping off of beans seedlings; at a concentration of 0,0625% decreases post emergence damping off in cucumber. In biochemical analyzes found an increase in the enzymatic activity of peroxidases and β-1,3-glucanases on beans, and glucanases on cucumber, and increased enzyme activity of peroxidases on beet, showing action in resistance induction at damping off.

Reator de leito estruturado com recirculação submetido à aeração intermitente no tratamento de esgoto sanitário

The release of nitrogen compounds in water bodies can result in many environmental problems, so treat wastewater, such as sewage in order to remove not only organic matter but also nitrogen has been studied a few decades. From the above, the objective of this study was to evaluate the performance of a structured bed reactor, continuous flow, with recirculation, in removing organic matter and nitrogen present in wastewater under different cycles of intermittent aeration (AI) and to evaluate the influence of these cycles in the development of nitrifying bacteria (Oxidizing Bacteria Ammonia - BOA and Bacteria Oxidizing Nitrite - BON) and denitrifying (DESN) adhered (Support Material - MS) and suspension (Effluent - EF and sludge - LD). The reactor used has usable volume of 9.4 L. As support materials (MS) polyurethane foam was used, cut and fixed in PVC rods. 3 were worked aeration phases (AE) and non-aeration (AN) at different stage: Stage 1 (4 h EA / AN 2H); Stage 2 (2H EA / AN 1 h) and Phase 3 (2H EA / AN 2 h). During all hydraulic detention time phases was kept at 16 h and the effluent recirculated at a rate of 3 times the inflow. Were analyzed: pH, total alkalinity, temperature, chemical oxygen demand (COD), Biochemical Oxygen Demand (BOD), nitrogen Kjeldhl Total (NKT), ammonia-N-N-NH4+, nitrito-N-NO2+andnitrato-NO3-. The concentration of BOA, BON and DESN was determined using the number More Provável.gSSV-1 (NMP.gSSV-1). In phase 1 the percentage removal NTK N-NH4+ and NT was 76±10%, 70±21% and 67±10% respectively. In Phase 2 80±15% of removel NKT, 86±15% of N-NH4+ e 68±9% of removel NT e na Fase 3 de 58±20%, 72±28% and 41±6% of NKT, N-NH4+ of NT, respectively. The denitrification efficiency in stage 3 was over 70%, indicating that occurred in the reactor the process of simultaneous nitrification and denitrification (NDS). DQOT the removal percentages were 88 ± 4% in Phase 1, 94 ± 7 in Phase 2 and 90± 11% in Phase 3. The multivariate ANOVA applied to NMP.gSSV-1, it indicated that there was significant (F: 20,2, p <0,01) between the analyzed concentration of organisms AI in different cycles, but the differences between NMP.gSSV-1 depends not only isolated factors but of which means, and phase groups being analysis. From the results it is concluded that the working system is efficient in terms of nitrogen removal and organic matter, and that the stage with the highest availability of Dissolved Oxygen (DO) and C/N ratio (Step 2), was the one obtained the lower concentrations of organic matter effluents and N-NH4+. Hinted that there was a significant difference between the concentration (NMP.100mL-1) of the analyzed organizations (BOA, BON and DESN), but this difference does not depend on factors alone but of which means (MS, EF or LD), stages (1, 2 or 3) and groups (BOA, BON and DESN) is being considered.

Óleo de carpa (cyprinus carpio): obtenção, refino e produção de concentração de ácidos graxos poliinsaturados

O objetivo do presente trabalho foi analisar as características e os perfis lipídicos dos óleos brutos e refinados de rejeitos de carpa comum (Cyprinus carpio), obtidos através dos processos de ensilagem ácida e termomecânico de farinha de pescado. Também foram realizados o estudo da reação de alcoólise química do óleo de carpa e obtenção de concentrados de concentrados de ácidos graxos poliinsaturados (PUFAs), através da reação de complexação com uréia. O rendimento obtido em óleo bruto para ambos os processos foi em torno de 85% em relação ao óleo presente nas vísceras de carpa. Os óleos brutos obtidos através do processo de ensilagem e do processo termomecânico apresentaram diferenças significantivas (P < 0,05) para ácidos graxos livres, índice de peróxido, valor do ácido tiobarbitúrico e cor Lovibond. Entretanto, os óleos refinados obtidos por ambos os processos não apresentaram diferença significativa para a cor Lovibond, ácidos graxos livres e valores do ácido tiobarbitúrico. Os principais ácidos graxos identificados nos óleos bruto, branqueado e refinado de vísceras de carpa foram oléico, palmítico, palmitoléico, linoléico e linolênico constituindo aproximadamente 69,6% dos ácidos graxo totais do óleo refinado. A relação ω3/ω6 foi de aproximadamente 1,05 para o óleo refinado. Assim, o óleo refinado das vísceras de carpa pode ser considerado uma rica fonte de ácidos graxos essenciais do grupo ω3 e ω6. No estudo da reação de alcoólise química e obtenção dos concentrados de ácidos graxos poliinsaturados, foi realizada a comparação de três tratamentos para reação de alcoólise variando-se a concentração molar óleo:álcool (1:21, 1:27 e 1:39). Os tratamentos apresentaram diferenças significativas para as respostas rendimento em massa de ácidos graxos livres e índice de acidez. O maior rendimento para a reação de alcoólise foi utilizando a concentração molar de 1:39 (óleo:álcool). Na fração não complexada com uréia obteve-se aumento percentual de ácidos graxos insaturados e poliinsaturados de 31,9%, redução de saturados de 75%, e aumento do conteúdo dos ácidos graxos eicosapentaenóico e docosahexaenóico (EPA+DHA) de 85,3%. A fração não complexada com uréia pode ser considerada uma rica fonte de ácidos graxos poliinsaturados e insaturados com um total de 88,9% desses ácidos graxos.

Medidas de concentração de radônio proveniente de argamassas de cimento portland, gesso e fosfogesso

Portland cement being very common construction material has in its composition the natural gypsum. To decrease the costs of manufacturing, the cement industry is substituting the gypsum in its composition by small quantities of phosphogypsum, which is the residue generated by the production of fertilizers and consists essentially of calcium dihydrate and some impurities, such as fluoride, metals in general, and radionuclides. Currently, tons of phosphogypsum are stored in the open air near the fertilizer industries, causing contamination of the environment. The 226 Ra present in these materials, when undergoes radioactive decay, produces the 222Rn gas. This radioactive gas, when inhaled together with its decay products deposited in the lungs, produces the exposure to radiation and can be a potential cause of lung cancer. Thus, the objective of this study was to measure the concentration levels of 222Rn from cylindrical samples of Portland cement, gypsum and phosphogypsum mortar from the state of Paraná, as well as characterizer the material and estimate the radon concentration in an environment of hypothetical dwelling with walls covered by such materials. Experimental setup of 222Rn activity measurements was based on AlphaGUARD detector (Saphymo GmbH). The qualitative and quantitative analysis was performed by gamma spectrometry and EDXRF with Au and Ag targets tubes (AMPTEK), and Mo target (ARTAX) and mechanical testing with x- ray equipment (Gilardoni) and the mechanical press (EMIC). Obtained average values of radon activity from studied materials in the air of containers were of 854 ± 23 Bq/m3, 60,0 ± 7,2 Bq/m3 e 52,9 ± 5,4 Bq/m3 for Portland cement, gypsum and phosphogypsum mortar, respectively. These results extrapolated into the volume of hypothetical dwelling of 36 m3 with the walls covered by such materials were of 3366 ± 91 Bq/m3, 237 ± 28 Bq/m3 e 208 ± 21 Bq/m3for Portland cement, gypsum and phosphogypsum mortar, respectively. Considering the limit of 300 Bq/m3 established by the ICRP, it could be concluded that the use of Portland cement plaster in dwellings is not secure and requires some specific mitigation procedure. Using the results of gamma spectrometry there were calculated the values of radium equivalent activity concentrations (Raeq) for Portland cement, gypsum and phosphogypsum mortar, which were obtained equal to 78,2 ± 0,9 Bq/kg; 58,2 ± 0,9 Bq/kg e 68,2 ± 0,9 Bq/kg, respectively. All values of radium equivalent activity concentrations for studied samples are below the maximum level of 370 Bq/kg. The qualitative and quantitative analysis of EDXRF spectra obtained with studied mortar samples allowed to evaluate quantitate and the elements that constitute the material such as Ca, S, Fe, and others.

CRESCIMENTO INICIAL E CONCENTRAÇÃO DE NUTRIENTES EM CLONES DE Eucalyptus urophylla x Eucalyptus grandis SOB INTERFERÊNCIA DE PLANTAS DANINHAS

A presença de plantas daninhas em plantios de eucalipto, especialmente nos dois primeiros anos, pode acarretar grandes prejuízos à produtividade, pois reduz a eficiência de aproveitamento dos recursos de crescimento pela cultura. Assim, objetivou-se com este trabalho avaliar os efeitos da interferência de plantas daninhas sobre o crescimento inicial de dois clones de Eucalyptus urophylla x Eucalyptus grandis e a concentração foliar de nutrientes na cultura e nas plantas daninhas. O experimento foi instalado em esquema fatorial 2 x 5 + 7, sendo dois clones de híbridos de Eucalyptus urophylla x Eucalyptus grandis, identificados como CNB001 e CNB016, em competição com cinco plantas daninhas Urochloa decumbens (capim-braquiária), Ipomoea nil (corda-de-viola), Commelina diffusa (trapoeraba), Spermacoce latifolia (erva-quente) e Panicum maximum (capim-colonião). Adicionalmente, foram cultivados os dois clones de eucalipto e as cinco plantas daninhas em monocultivo como padrão de comparação, no delineamento inteiramente casualizado, com quatro repetições. Foram avaliados eucalipto através da altura de plantas, o diâmetro do coleto, o número de ramos, a área foliar, a matéria seca e, o teor foliar de nutrientes do eucalipto, bem como o teor de nutrientes nas folhas das plantas daninhas. O clone CNB001 apresentou crescimento inicial superior ao clone CNB016, no entanto, livre da interferência de plantas daninhas, verificaram-se teores foliares semelhantes para a maioria dos nutrientes em ambos os genótipos. O clone CNB016 mostrou maior sensibilidade à interferência negativa das plantas daninhas que o clone CNB001, sendo seu crescimento inicial mais afetado por Ipomoea nil e a concentração de nutrientes reduzida pelas espécies Panicum maximum, Urochloa decumbens e Commelina diffusa. Panicum maximum apresentou maior interferência com o clone CNB001, enquanto Ipomoea nil pouco influenciou o crescimento e o teor de nutrientes deste híbrido. As plantas daninhas apresentaram elevada capacidade de extrair nutrientes do solo, mesmo em convivência com os clones de Eucalyptus urophylla x Eucalyptus grandis. A interferência imposta à cultura é dependente da espécie infestante e do genótipo de eucalipto.

Avaliação da concentração de retinol em fígados frescos e congelados de frangos das linhagens Cobb e Ross

The intake of adequate quantities of food, including those rich in vitamins, is necessary for a healthy life. The lack of vitamin A has been characterized as a public health problem in developing countries, however, a high intake of vitamin A can result in toxic and teratogenics effects. High concentrations of vitamin A have been observed in the livers of animals. The objective of this study was to assess the levels of retinol in chicken livers and verify the effect of frozen storage on these levels. 64 livers from two chicken strains, Cobb and Ross, were used, came from four different farms. We examined 32 livers from each strain, 8 samples from each farm. Liver sample were homogenized individually, then 4 aliquots were taken from each sample. One of aliquots was analyzed immediately after slaughter (T0), the others were analyzed after 30, 60 and 90 days of storage at -18oC (T30, T60 and T90, respectively). Retinol dosage in the liver was determined by High Performance Liquid Chromatography (HPLC). The levels of retinol varied significantly according to the strain. The mean retinol value in the fresh samples was 6678.0 ± 1337.7 and 8324.1 ± 1158.5 µg/100g in the Cobb and Ross strain, respectively. Values of 4258 ± 918.7 ± 1391.7 and 4650.5 ± 1391.7 μg/100g were found after 90 days of storage for Cobb and Ross strain, respectively. The liver freezing caused a significant reduction in their levels of retinol, causing a loss of up to 44% with respect to fresh livers. The reduction in retinol levels occurred from 30 days of storage. Even with the losses from the frozen, the ingestion of a typical portion of 100 g of liver, regardless the chicken strain analyzed, surpass all recommendations of consumption and the maximum tolerable intake of vitamin A (3000 μg/day) for adults

Desenvolvimento de método empregando DLLME para extração e pré-concentração de ferro em amostras de água com diferentes salinidades

Neste trabalho foi desenvolvido e validado um método analítico para a determinação de Fe total em amostras de água com diferentes salinidades empregando a microextração líquido-líquido dispersiva (DLLME), seguido de determinação espectrofotométrica na região do ultravioleta-visível. Nas extrações por DLLME, o Fe foi complexado com pirrolidina ditiocarbamato de amônio (APDC) e extraído após a injeção de uma mistura composta de 1,2-diclorobenzeno (solvente extrator) e etanol (solvente dispersor). Os fatores que influenciam a eficiência de extração (tipo e volume de solvente extrator e dispersor, concentração de APDC e tempo de extração) e subsequente determinação espectrofotométrica foram otimizados. Após otimização das condições, a curva de calibração foi linear entre 0,02 e 2,0 mg L-1 (r 2 = 0,9998) e os limites de detecção e quantificação do método foram de 6,1 e 18,3 µg L-1 , respectivamente. O fator de pré-concentração foi de 3,3. A exatidão foi avaliada em termos de recuperação, com variação entre 90 e 102% e desvio padrão relativo (RSD) inferiores a 12%. A exatidão do método também foi avaliada utilizando um material de referência certificado (SRM 1643e, Elementos Traço em Água), o qual apresentou concordância de 104% e RSD de 5%. O método foi aplicado para amostras de água com diferentes salinidades, como água mineral, estuarina e de alto mar. Além disso, o método proposto apresentou concordância da ordem de 95%, quando comparado com o método 1,10-fenantrolina para a determinação de Fe total.

Efeito da suplementação materna com palmitato de retinila sobre a concentração de retinol no colostro em condições de jejum e pós-prandial.

Vitamin A is important in many essential body processes and its deficiency results in serious consequences for human health. Breast milk is the only source of this vitamin for children that are exclusively breastfed. Analysis of vitamin A in mother s milk is important because its concentration is related to maternal vitamin A status and to its ingestion by the mother during pregnancy. The aim of the present study was to assess the effect of maternal supplementation with retynil palmitate on the concentration of colostrum retinol under fasting and postprandial conditions. A total of 149 nursing mothers were recruited at the Januário Cicco Maternity School (Natal, Brazil) and allocated to two groups: Comparison (n = 69) and Test (n = 80). Blood and colostrum (in fasting and postprandial conditions) samples were collected up to 24hs after delivery. Serum retinol and colostrum levels were analyzed by high-performance liquid chromatography. The serum retinol level of 41.6 ± 12.7μg/dL (mean ± standard deviation) indicates adequate biochemical nutritional status. Colostrum retinol level was not influenced by serum retinol levels under any of the conditions established. In the colostrum, the retinol concentration in the unsupplemented test group was 67.3 ± 37.7 μg/dL under fasting and 80.3 ± 35.1 μg/dL under postprandial conditions (p<0.05), showing an increase of 19.3%. In the supplemented test group the values were 102.6 ± 57.3 μg/dL and 133.4 ± 78.3 μg/dL under fasting and postprandial, respectively (p<0.05), representing an increase of 30%. Considering that under fasting conditions most of the vitamin A transported to the milk originates in the retinol binding protein (RBP), the postprandial increase in colostrum retinol suggests a different transport mechanism of retinol to maternal milk from that performed by RBP. This situation becomes more evident under supplementation conditions.

Resposta de Chromobacterium Violaceum cultivada em alta concentração de ferro

The Chromobacterium violaceum is a β-proteobacterium Gram-negative widely found in tropical and subtropical regions, whose genome was sequenced in 2003 showing great metabolic versatility and biotechnological and pharmaceutical potential. Given the large number of ORFs related to iron metabolism described in the genome of C. violaceum, the importance of this metal for various biological processes and due to lack of data about the consequences of excess of iron in free-living organisms, it is important to study the response mechanism of this bacterium in a culture filled with iron. Previous work showed that C. violaceum is resistant to high concentrations of this metal, but has not yet been described the mechanism which is used to this survival. Thus, to elucidate the response of C. violaceum cultured in high concentrations of iron and expecting to obtain candidate genes for use in bioremediation processes, this study used a shotgun proteomics approach and systems biology to assess the response of C. violaceum grown in the presence and absence of 9 mM of iron. The analysis identified 531 proteins, being 71 exclusively expressed by the bacteria grown in the presence of the metal and 100 just in the control condition. The increase in expression of proteins related to the TCA cycle possibly represents a metabolic reprogramming of the bacteria caused by high concentration of iron in the medium. Moreover, we observed an increase in the activity assay of superoxide dismutase and catalase as well as in Total Antioxidant Activity assay, suggesting that the metal is inducing oxidative stress in C. violaceum that increases the levels of violacein and antioxidant enzymes to better adapt to the emerging conditions. Are also part of the adaptive response changes in expression of proteins related to transport, including iron, as well as an increased expression of proteins related to chemotaxis response, which would lead the bacteria to change the direction of its movement away from the metal. Systems Biology results, also suggest a metabolic reprogramming with mechanisms coordinated by bottleneck proteins involved in transcription (GreA), energy metabolism (Rpe and TpiA) and methylation (AhcY)