Increasing consumer demand for new products with high nutritional qualities has spurred a search for new alternatives to food preservation. Pulsed electric field (PEF) is an emerging technology for non thermal food pasteurisation. Using this technology, enzymes, pathogenic and spoilage microorganisms can be inactivated without affecting the colour, flavour, and nutrients of the food. PEF treatment may be provided by applying pulsed electric field to a food product in a treatment zone between two electrodes at ambient , or slightly above ambient temperature. Exposure of microbial cells to the electric field induces a transmembrane potential in the cell membrane, which results in electroporation (the permeabilization of the membranes of cells and organelles) and/or electrofusion (the connection of two separate membranes into one) of the cells. An innovative pulsed electric field (PEF) unit was designed and constructed in the University of Auckland using modern IGBT technology. The system consists of main equipments, the high voltage pulse generator and the treatment chambers. The main focus of this work was to design an innovative PEF treatment chamber that provide uniform distribution of electric field, minimum increase in liquid temperature, minimum fouling of electrodes and an energy efficient system. Four multi pass treatment chambers were designed consisting of two stainless steel mesh electrodes in each chamber, with the treated fluid flowing through the openings of the mesh electrodes. The two electrodes are electrically isolated from each other by an insulator element designed to form a small orifice where most of the electric field is concentrated. Dielectric breakdown inside the chambers was prevented by removing the electrodes far from the narrow gap. The effect of the chambers different geometries on the PEF process in terms of electric parameters and microbial inactivation were investigated. Electric field intensity in the range of (17-43 kV/cm) was applied with square bipolar pulses of 1.7 µs duration. The effect of PEF treatment on the inactivation of gram-negative Escherichia coli ATCC 25922 suspended in simulated milk ultra-filtrate (SMUF) of 100%, 66.67% and 50% concentration was investigated. Treatments with the same electrical power input but higher electric field strengths provided larger degree of killing. The inactivation rate of E coli was significantly increased with increasing the electric field strength, treatment time and processing temperature. Morphological changes on E coli as a result of PEF treatment were studied under transmission electron microscopy (TEM). Significant morphological changes on E coli after PEF treatment were observed. The TEM studies suggested that the microbial inactivation was a consequence of electroporation and electrofusion mechanisms. Kinetic analysis of microbial inactivation due to PEF and thermal treatment of E coli suspended in SUMF were also studied. Comparison between measured (experimental) and predicted (theoretical) variation of E coli concentration with time following the PEF treatment was discussed, taking into consideration the recirculation mode of the PEF treatment. The treated liquid was circulated more than once through the treatment chamber to provide higher microbial inactivation. Arrhenius constants and activation energies of E coli inactivation using combined PEF and thermal treatment were calculated and generalized correlation for the inactivation rate constant as a function of electric field intensity and treatment temperature was developed. / Fonterra Research Institute (NZ) and the Foundation for Research Science and Technology (NZ)

Increasing consumer demand for new products with high nutritional qualities has spurred a search for new alternatives to food preservation. Pulsed electric field (PEF) is an emerging technology for non thermal food pasteurisation. Using this technology, enzymes, pathogenic and spoilage microorganisms can be inactivated without affecting the colour, flavour, and nutrients of the food. PEF treatment may be provided by applying pulsed electric field to a food product in a treatment zone between two electrodes at ambient , or slightly above ambient temperature. Exposure of microbial cells to the electric field induces a transmembrane potential in the cell membrane, which results in electroporation (the permeabilization of the membranes of cells and organelles) and/or electrofusion (the connection of two separate membranes into one) of the cells. An innovative pulsed electric field (PEF) unit was designed and constructed in the University of Auckland using modern IGBT technology. The system consists of main equipments, the high voltage pulse generator and the treatment chambers. The main focus of this work was to design an innovative PEF treatment chamber that provide uniform distribution of electric field, minimum increase in liquid temperature, minimum fouling of electrodes and an energy efficient system. Four multi pass treatment chambers were designed consisting of two stainless steel mesh electrodes in each chamber, with the treated fluid flowing through the openings of the mesh electrodes. The two electrodes are electrically isolated from each other by an insulator element designed to form a small orifice where most of the electric field is concentrated. Dielectric breakdown inside the chambers was prevented by removing the electrodes far from the narrow gap. The effect of the chambers different geometries on the PEF process in terms of electric parameters and microbial inactivation were investigated. Electric field intensity in the range of (17-43 kV/cm) was applied with square bipolar pulses of 1.7 µs duration. The effect of PEF treatment on the inactivation of gram-negative Escherichia coli ATCC 25922 suspended in simulated milk ultra-filtrate (SMUF) of 100%, 66.67% and 50% concentration was investigated. Treatments with the same electrical power input but higher electric field strengths provided larger degree of killing. The inactivation rate of E coli was significantly increased with increasing the electric field strength, treatment time and processing temperature. Morphological changes on E coli as a result of PEF treatment were studied under transmission electron microscopy (TEM). Significant morphological changes on E coli after PEF treatment were observed. The TEM studies suggested that the microbial inactivation was a consequence of electroporation and electrofusion mechanisms. Kinetic analysis of microbial inactivation due to PEF and thermal treatment of E coli suspended in SUMF were also studied. Comparison between measured (experimental) and predicted (theoretical) variation of E coli concentration with time following the PEF treatment was discussed, taking into consideration the recirculation mode of the PEF treatment. The treated liquid was circulated more than once through the treatment chamber to provide higher microbial inactivation. Arrhenius constants and activation energies of E coli inactivation using combined PEF and thermal treatment were calculated and generalized correlation for the inactivation rate constant as a function of electric field intensity and treatment temperature was developed. / Fonterra Research Institute (NZ) and the Foundation for Research Science and Technology (NZ)

Increasing consumer demand for new products with high nutritional qualities has spurred a search for new alternatives to food preservation. Pulsed electric field (PEF) is an emerging technology for non thermal food pasteurisation. Using this technology, enzymes, pathogenic and spoilage microorganisms can be inactivated without affecting the colour, flavour, and nutrients of the food. PEF treatment may be provided by applying pulsed electric field to a food product in a treatment zone between two electrodes at ambient , or slightly above ambient temperature. Exposure of microbial cells to the electric field induces a transmembrane potential in the cell membrane, which results in electroporation (the permeabilization of the membranes of cells and organelles) and/or electrofusion (the connection of two separate membranes into one) of the cells. An innovative pulsed electric field (PEF) unit was designed and constructed in the University of Auckland using modern IGBT technology. The system consists of main equipments, the high voltage pulse generator and the treatment chambers. The main focus of this work was to design an innovative PEF treatment chamber that provide uniform distribution of electric field, minimum increase in liquid temperature, minimum fouling of electrodes and an energy efficient system. Four multi pass treatment chambers were designed consisting of two stainless steel mesh electrodes in each chamber, with the treated fluid flowing through the openings of the mesh electrodes. The two electrodes are electrically isolated from each other by an insulator element designed to form a small orifice where most of the electric field is concentrated. Dielectric breakdown inside the chambers was prevented by removing the electrodes far from the narrow gap. The effect of the chambers different geometries on the PEF process in terms of electric parameters and microbial inactivation were investigated. Electric field intensity in the range of (17-43 kV/cm) was applied with square bipolar pulses of 1.7 µs duration. The effect of PEF treatment on the inactivation of gram-negative Escherichia coli ATCC 25922 suspended in simulated milk ultra-filtrate (SMUF) of 100%, 66.67% and 50% concentration was investigated. Treatments with the same electrical power input but higher electric field strengths provided larger degree of killing. The inactivation rate of E coli was significantly increased with increasing the electric field strength, treatment time and processing temperature. Morphological changes on E coli as a result of PEF treatment were studied under transmission electron microscopy (TEM). Significant morphological changes on E coli after PEF treatment were observed. The TEM studies suggested that the microbial inactivation was a consequence of electroporation and electrofusion mechanisms. Kinetic analysis of microbial inactivation due to PEF and thermal treatment of E coli suspended in SUMF were also studied. Comparison between measured (experimental) and predicted (theoretical) variation of E coli concentration with time following the PEF treatment was discussed, taking into consideration the recirculation mode of the PEF treatment. The treated liquid was circulated more than once through the treatment chamber to provide higher microbial inactivation. Arrhenius constants and activation energies of E coli inactivation using combined PEF and thermal treatment were calculated and generalized correlation for the inactivation rate constant as a function of electric field intensity and treatment temperature was developed. / Fonterra Research Institute (NZ) and the Foundation for Research Science and Technology (NZ)

Submitted by Ligia Souza (ligia@ufscar.br) on 2017-09-21T14:27:46Z No. of bitstreams: 1 TeseMCG.pdf: 3682312 bytes, checksum: fd17a7f18e76507395baaaf315348805 (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2017-09-25T12:32:38Z (GMT) No. of bitstreams: 1 TeseMCG.pdf: 3682312 bytes, checksum: fd17a7f18e76507395baaaf315348805 (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2017-09-25T12:32:44Z (GMT) No. of bitstreams: 1 TeseMCG.pdf: 3682312 bytes, checksum: fd17a7f18e76507395baaaf315348805 (MD5) / Made available in DSpace on 2017-09-25T12:37:57Z (GMT). No. of bitstreams: 1 TeseMCG.pdf: 3682312 bytes, checksum: fd17a7f18e76507395baaaf315348805 (MD5) Previous issue date: 2017-07-31 / Outra / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Infectious pneumonia is a major cause of morbidity/mortality, mainly due to the increasing rate of microorganisms resistant to antibiotics. Photodynamic Inactivation (PDI) is emerging as a promising approach, as effects are based on oxidative stress, preventing the emergence of resistant microorganism strains. In previous studies, the in vitro inactivation of Streptococcus pneumoniae using indocyanine green (ICG) and infrared light source was successful, and achieved reduction of 5 log10 colony-forming units (CFU/mL) with concentration as low as 10 μM ICG. In the present study, a proof-of-principle protocol was designed to treat lung infections by PDI using extracorporeal illumination with a 780 nm laser device and ICG as photosensitizer. In a first row of experiments, hairless mice were infected with S. pneumoniae and PDI was performed two days after infection. For control groups, CFU recovery ranged between 103-104 CFU/mL/mouse. For PDT group, however, no bacteria were recovered in 80% of the animals. Animal survival was evaluated separately over 50 days. No deaths occurred in PDT group, whereas 60% of the control did not survive. Lung injury analyses were performed in BALB/c mice, the bacteria reduction were 2 and 4 log10 in 2 mice (5 in total) and the wet-to-dry ratio showed that PDI did not increase the edema in lungs. The bronchoalveolar lavage data indicated a larger absolute number of cells (mononuclear and polymorphonuclear) in the PDI group in contrast to control group, meaning that the technique could increase the immune system response. In vitro results showed the irradiated ICG could generate aggregates or photoproducts that help PDI to inactivate the bacteria. Our results indicate that extracorporeal PDI has potential for pneumonia treatment, and pulmonary decontamination with PDI may be used as a single therapy or as an antibiotics adjuvant. / A pneumonia infecciosa é uma das principais causas de morbidade/mortalidade no mundo, principalmente devido à taxa crescente de micro-organismos resistentes a antibióticos. A inativação fotodinâmica (IFD) está emergindo como uma abordagem promissora, cujos efeitos são baseados no estresse oxidativo, impedindo o surgimento de cepas de micro-organismos resistentes. Em estudos anteriores, a inativação in vitro de Streptococcus pneumoniae utilizando indocianina verde (ICV) e fonte de luz de infravermelho foi efetiva, inativando 5 log10 (UFC/mL) com apenas 10 μM ICV. Neste estudo, foi avaliado protocolo de prova de princípio para tratar infecções pulmonares por IFD usando irradiação extracorpórea com dispositivo a laser emitindo em 780 nm e ICV como fotossensibilizador (FS). Em uma primeira série de experimentos, camundongos hairless SKH-1 foram infectados com S. pneumoniae e a IFD foi realizada dois dias após a infecção. Para os grupos de controles, a recuperação de UFC variou entre 103-104 UFC/mL/animal. Para o grupo IFD, no entanto, nenhuma bactéria foi recuperada em 80% dos animais. A sobrevivência animal foi avaliada durante 50 dias. Não ocorreram mortes no grupo IFD, enquanto 60% do grupo controle foi a óbito. Foram realizadas análises de lesões pulmonares em camundongos BALB/c, onde a redução bacteriana foi de 2 e 4 log10 em dois animais (total 5) em comparação com o grupo controle, e a proporção de peso úmido e seco mostrou que a IFD não aumentou o edema nos pulmões. Os dados de lavagem broncoalveolar indicaram um aumento no número absoluto de células (mononucleares e polimorfonucleares) no grupo IFD, o que indica que a técnica pode aumentar a resposta do sistema imunológico. Os resultados in vitro mostraram que a ICV irradiada pode gerar agregados ou fotoprodutos que auxiliam a IFD a inativar a bactéria. Os resultados deste estudo indicam que a IFD com irradiação extracorpórea tem potencial para o tratamento de pneumonia, e a descontaminação pulmonar com IFD pode ser usada como terapia ou como adjuvante aos antibióticos. / CAPES: 99999.003154/2015-07

Made available in DSpace on 2014-06-11T19:32:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-25Bitstream added on 2014-06-13T20:24:21Z : No. of bitstreams: 1 pavezzi_fc_dr_rcla.pdf: 679532 bytes, checksum: a99630bf198f33da999b112631255c08 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A glucoamilase é uma enzima hidrolítica que catalisa a liberação sucessiva de β-D-glicose a partir do amido e oligossacarídeos relacionados. Neste trabalho foram estudadas as glucoamilases de Aspergillus awamori expressas em levedura Saccharomyces cerevisiae. Foram utilizadas duas linhagens alteradas denominadas M1 e M2, e uma linhagem selvagem (WT), utilizada como parâmetros na comparação dos resultados. As enzimas foram produzidas em fermentação submersa, e amidos de diferentes origens vegetais foram utilizados como uma fonte extra de carbono na produção das enzimas. O melhor substrato para a produção da glucoamilase selvagem e da mutante M2 foi o amido de batata com 8,2 e 6,6 U/mL, respectivamente. Para a linhagem M1 foi o amido de mandioca com atividade enzimática de 5,9 U/mL. O amido de milho mostrou ser um substrato menos indicado para a produção destas enzimas. Para a purificação foi preparada uma coluna de afinidade com resina sepharoseTM 6B epóxi ativada ligada a acarbose, onde diferentes concentrações do ligante foram avaliadas. A coluna apresentou boa eficiência no processo de purificação conforme análise por eletroforese SDS-PAGE, com massas moleculares estimada em 100 kDa. A temperatura ótima de atividade das enzimas M1 e M2 foi 65 °C, enquanto que a selvagem teve sua atividade máxima em 60 °C. O pH ótimo de atuação das enzimas foi 4,5. As glucoamilases mutantes apresentaram maior termoestabilidade que a glucoamilase selvagem durante o processo de termoinativação, destacando principalmente a glucoamilase M2. A meia vida a 70 °C foi de 8,1 minutos para a enzima mutante M2, 4,1 minutos para a M1 e 3,0 minutos para a enzima selvagem. A energia de ativação para a desnaturação (Ead) foi de 252,9 e 262,8 KJ mol-1 para as enzimas M1 e M2 respectivamente, e de 234,3 KJ mol-1 para a selvagem. A maior energia dos mutantes indica maior resistência... / Glucoamylase is a hydrolytic enzyme that catalyzes the consecutive liberation of β-D-glucose from starch and related oligosaccharides. In this work glucoamylases from Aspergillus awamori expressed in the yeast Saccharomyces cerevisiae were studied. Two mutant strains, denominated M1 and M2, were used and one wild strain (WS) was used as parameter to compare the results. The enzymes were produced in submerged fermentation and starches from different botanical origins were used as extra carbon source for enzyme production. The best substrate for the production of wild glucoamylase and of mutant M2 was potato starch with 8.2 and 6.6 U/mL, respectively. For strain M1 the best substrate was cassava starch with enzymatic activity of 5.9 U/mL. Corn starch revealed to be a less indicated starch for the production of these enzymes. For purification, an affinity column was prepared with activated SepharoseTM 6B epoxy linked to acarbose, and different concentrations of ligand were evaluated. The column exhibited good efficiency during the purification process according to SDS-PAGE analysis, with molecular masses estimated in 100 kDa. Optimum temperature for activities of M1 and M2 enzymes was 65°C, while the wild one exhibited maximum activity at 60°C. Optimum pH for enzyme action was 4.5. Mutant glucoamylases presented higher thermostability than wild glucoamylase during the thermoinactivation process, with M2 standing out. Half life at 70°C was of 8.1 minutes for mutant enzyme M2, 4.1 minutes for M1 and 3.0 minutes for wild enzyme. Activation energy for denaturation (Ead) was 252.9 and 262.8 KJ mol-1 for enzymes M1 and M2 respectively, and 234.3 KJ mol-1 for the wild one. The higher energy of the mutants indicates higher resistance of the protein structure, since more energy is required for the molecule to enter a transition and unfolding state. Thermodynamic parameter ΔG was higher... (Complete abstract click electronic access below)

Voltage-gated Cav2.1 (P/Q-type) and Cav2.2 (N-type) channels are two closely related calcium channels that play indispensable roles in signal transduction pathways by regulating neurotransmitter release. Despite having highly conserved amino acid sequences, they are differentially modulated by calmodulin, which mediate two important feedback mechanisms known as Ca2+-dependent inactivation (CDI) and Ca2+-dependent facilitation (CDF). These dual regulatory mechanisms contribute to synaptic plasticity, but only CDI is observed in Cav2.2 channel, while both CDI and CDF are present in Cav2.1 channel. Previously, it was hypothesized that the lack of CDF in Cav2.2 channel is due to the pre-IQ-IQ domain of the channel’s lower binding affinity for calmodulin compared to that of Cav2.1 channel. Now that the EF-hand domain of calcium channels is identified as one of the two minimally required molecular determinants that are responsible for supporting CDF in Cav2.1 channel and preventing CDF in Cav2.2 channel, it was necessary to determine the role of EF-hand domain in calmodulin binding of Cav2.1 and Cav2.2 channels. Using pull-down binding assays, this study finds that the EF-hand domain enhances calmodulin binding to the proximal C-terminal domain of Cav2.2 channel, which suggests that the lack of CDF in Cav2.2 does not result from the channel’s weak interaction with CaM, but from the EF-pre-IQ-IQ domain of the channel’s inability to allow calmodulin from fully exerting its effects.

O alho (Allium sativum L.) originário das zonas temperadas da Ásia Central é uma planta herbácea da família Alliaceae; possui como principais constituintes funcionais a alicina e a inulina. A alicina é um componente ativo e responsável pelo cheiro característico e atividade antimicrobiana, enquanto que a inulina é classificada como prebiótico e como fibra alimentar solúvel por ser resistente à digestão na parte superior do trato intestinal. Neste trabalho foram estudadas as propriedades físico-químicas do alho in natura, além do processo de branqueamento em água e vapor e a desidratação por ar quente e liofilização no alho. Os bulbos de alho foram limpos e selecionados considerando a ausência de injúrias visuais e infecções, bem como a uniformidade de tamanho e cor. O alho in natura apresentou umidade de 64,15±0,09 %, atividade de água de 0,986±0,001, sólidos solúveis de 36,00±0,85 °Brix e pH de 6,41±0,008. As concentrações de inulina, glicose e frutose foram de 56,62±0,89, 2,37±0,03 e 2,23±0,05 g/100g de matéria seca respectivamente. Os bulbilhos do alho foram descascados e cortados em rodelas com diâmetros de 15±2,40 mm e espessuras de 1±0,35 mm. A seguir as amostras passaram pelo processo de branqueamento em água previamente aquecido a 80 e 90 °C e em vapor a 100 °C à pressão atmosférica. Foram empregados tempos de 1, 2, 4, 6, 8 e 10 minutos em ambos os casos. Nesta etapa verificou-se o efeito do tempo e temperatura de branqueamento sobre a atividade das enzimas peroxidase, polifenoloxidase e inulinase e, mediram-se os parâmetros de cor L*, a* e b*, avaliando-se a cinética de inativação dessas enzimas e das mudanças de cor. A análise das concentrações de inulina, glicose e frutose foram realizadas para as melhores condições de branqueamento utilizando Cromatografia Líquida de Alta Eficiência. A melhor condição de branqueamento foi obtida no vapor por 4 minutos, onde não se observaram mudanças na textura, com redução da atividade enzimática de 93,53 %, 92,15 % e 81,96 % para a peroxidase, polifenoloxidase e inulinase respectivamente. Nesta condição a concentração da inulina diminuiu 3,72 %, enquanto que a glicose aumentou 0,67 % e a frutose 0,55 %, quando comparadas ao alho in natura, devido à atividade residual da inulinase. Durante o branqueamento em água e vapor o parâmetro de cor L* aumentou com o tempo, tornando as amostras mais claras, enquanto que a* e b* diminuíram, obtendo-se rodelas mais esverdeadas e azuladas. Na secagem, as rodelas de alho sem ou com branqueamento em vapor por 4 minutos, foram levadas a um secador de ar forçado às temperaturas de 50, 60 e 70 °C durante 6 horas. Para a liofilização as rodelas foram previamente congeladas a -80 °C por 24 horas e dispostas em bandejas dentro de um liofilizador empregando pressão de 64 μmHg, onde permaneceram por tempo médio de 48 horas. Ao estudar a cinética de secagem em ar forçado usando os modelos propostos por Henderson-Pabis, Page e Newton constatou-se o aumento da constante da taxa de secagem com o aumento da temperatura e com o uso do branqueamento. Os valores de umidade e de atividade de água no equilíbrio, obtidos a partir da porção assintótica das curvas de secagem em condições dinâmicas, estiveram na faixa de 0,086 a 0,102 g/g de matéria seca e de 0,376 a 0,521 aw respectivamente, sendo estes menores com o aumento da temperatura e com emprego do branqueamento. Posteriormente as amostras desidratadas por ar quente e liofilização, foram moídas para medição dos parâmetros de cor, tamanho de partículas e determinação dos teores de inulina, glicose e frutose, temperatura de transição vítrea e observação da microestrutura por microscopia eletrônica de varredura. As amostras liofilizadas tiveram valores de L* significativamente maiores, e parâmetros de a* e b* menores que as amostras desidratadas em ar forçado, sendo as amostras liofilizadas mais claras, esverdeadas e azuladas, próxima à cor do alho in natura. Também foi encontrado a diminuição da concentração de inulina e aumento dos teores de glicose e frutose nas amostras desidratadas, indicando a possível hidrólise da inulina, podendo estar relacionada à atividade residual da enzima inulinase. Também foi observado nas amostras branqueadas menores concentração de inulina, glicose e frutose, que quando não branqueadas, devido à possível lixiviação na água. Nas amostras liofilizadas os diâmetros médios das partículas foram de 79,35 μm e 104,79 μm no alho sem e com branqueamento respectivamente, sendo menores que às desidratadas em ar quente que variaram de 119,28 μm à 302,04 μm, observando-se, por microscopia eletrônica, menores danos na superfície das amostras liofilizadas, pois este processo origina menor contração e conseqüentemente menor rugosidade nas amostras quando comparado com a secagem em ar. As temperaturas de transição vítrea do alho desidratado em pó variaram de 99,39±1,98 °C à 120,15±3,80 °C, sendo maior quanto menor o valor de atividade de água, confirmando o efeito plasticizante da água. / Garlic (Allium sativum L.) originated in the temperate zones of Central Asia and is a herbaceous plant of the Alliaceae family; its main functional components are allicin and inulin. Allicin is the active component responsible for the characteristic odor and anti-microbial activity, whereas inulin is classified as a prebiotic substance and as a soluble dietary fiber since it is resistant to digestion in the small intestine. The physicochemical properties of garlic in natura were studied in the present work, and also the processes of water and steam blanching and dehydration in hot air and by freeze-drying. The garlic cloves were cleaned and selected considering the absence of visual injuries and infections and also uniformity of size and color. The in natura garlic presented a moisture content of 64.15±0.09 %, water activity of 0.986±0.001, soluble solids of 36.00±0.85 °Brix and pH of 6.41±0.008. The inulin, glucose and fructose concentrations were 56.62±0.89, 2.37±0.03 and 2.23±0.05 g/100g of dry matter, respectively. The garlic cloves were peeled and cut into slices with diameters of 15±2.40 mm and thicknesses of 1±0.35 mm. The samples were submitted to blanching in water baths previously heated to 80 and 90 ºC and, steam at a temperature of 100 ºC at atmospheric pressure. Times of 1, 2, 4, 6, 8 and 10 minutes were employed for both water and steam blanching. At this stage the effects of blanching time and temperature on the activities of the enzymes peroxidase, polyphenoloxidase and inulinase were determined, and the color parameters of L*, a* and b* were measured, evaluating the kinetics of inactivation of these enzymes and the color changes. The concentrations of inulin, glucose and fructose were determined by high performance liquid chromatography in the samples treated with the best of the blanching conditions. The best blanching conditions were obtained using steam for 4 minutes. Under these conditions no changes in texture were observed, and the enzymatic activities were reduced by 93.53 %, 92.15 % and 81.96 % for peroxidase, polyphenoloxidase and inulinase, respectively. Under these conditions the inulin concentration decreased by 3.72 % and the glucose and fructose concentrations increased by 0.67 % and 0.55 %, respectively, when compared to the in natura garlic, due to the residual inulinase activity. The color parameter L* increased with time during both steam and water blanching, whereas a* and b* decreased, obtaining slices which were greener and bluer. For drying, the garlic slices, with or without steam blanching for 4 minutes, were placed in forced air dryers at 50, 60 and 70 ºC for 6 hours. For freeze drying, the slices were previously frozen at -80 ºC for 24 hours and then placed in trays in the freeze drier at a pressure of 64 μmHg, where they remained for a mean time of 48 hours. The models proposed by Henderson-Pabis, Page & Newton were used to study the drying kinetics, and it was shown that the drying rate constant increased with increase in temperature and with the use of blanching. The equilibrium values for moisture content and water activity (aw), obtained from the asymptotic part of the drying curves under dynamic conditions, were in the range from 0.086 to 0.102 g/g of dry matter and from 0.376 to 0.521 respectively, these values decreasing with increase in temperature and with the use of blanching. After drying the samples using both hot air and freeze-drying, they were ground for subsequent determination of the color parameters, particle size, inulin, glucose and fructose contents and glass transition temperatures, and for observation of the microstructure by scanning electronic microscopy. The freeze-dried samples showed significantly higher values for L* and lower values for a* and b* than the samples dried by forced air, and thus the freeze-dried samples were lighter in color and more greenish or bluish, closer to the color of the in natura garlic. All the dehydrated samples showed a decrease in the inulin contents and increase in the glucose and fructose contents, indicating a possible hydrolysis of the inulin which could be related to the residual activity of the enzyme inulinase. There were also lower inulin, glucose and fructose concentrations in the blanched samples as compared to the non-blanched samples, possibly due to leaching into the water. In the freeze-dried samples the mean particle diameters were 79.35 μm and 104.79 μm for the non-blanched and blanched samples, respectively, smaller than the values obtained in the forced air-dried samples, whose diameters varied from 119.28 μm to 302.04 μm. By way of scanning electronic microscopy, it was observed that the surface of the freeze-dried samples was less damaged than that of the forced air-dried samples, since freeze-drying results in less contraction and consequently less wrinkling than air drying. The glass transition temperatures of the dehydrated garlic powders varied from 99.39±1.98 °C to 120.15±3.80 °C, increasing with decrease in the water activity, confirming the plasticizing effect of water.

O hibisco (Hibiscus sabdariffa L.) além de possuir propriedades antioxidantes e antimicrobianas, é utilizado como planta medicinal e alimento funcional nos países tropicais. Através de Testes de Diluição em Sistema de Tubos Múltiplos determinou-se a Intensidade de Atividade de Inibição Bacteriana (IINIB/Bacteriostasia) e a Intensidade de Atividade de Inativação Bacteriana (IINAB/Bactericidia) de extrato alcoólico de dois acessos de hibisco, a saber: Palmares do Sul/RS e Porto Alegre/RS sobre inóculos padronizados de Enterococcus faecalis (ATCC 19433), Escherichia coli (ATCC 11229), Salmonella Enteritidis (ATCC 11076) e Staphylococcus aureus (ATCC 25923). Paralelamente, o teor de polifenóis totais e de antocianinas nos cálices e nos frutos com sementes do hibisco foi determinado. A atividade antimicrobiana do extrato alcoólico de cálices, em ambos os acessos, apresentou diferença positiva significativa quando relacionada ao extrato alcoólico dos frutos com sementes. Salmonella Enteritidis foi a bactéria mais sensível ao extrato alcoólico de cálices do hibisco enquanto a mais resistente foi Staphylococcus aureus, independente da variável acesso, considerando somente a estrutura vegetal. Em relação ao extrato alcoólico dos frutos com sementes, Escherichia coli demonstrou a maior sensibilidade e Staphylococcus aureus a maior resistência. Os valores de polifenóis totais e de antocianinas do extrato alcoólico de cálices apresentaram diferença significativa entre si e foram superiores ao extrato alcoólico dos frutos com sementes. Possivelmente existe uma relação direta entre a concentração de antocianina e a atividade antibacteriana em diferentes estruturas vegetais do hibisco. / The hibiscus (Hibiscus sabdariffa L.) has antioxidant and antimicrobial properties and it is utilized as functional food and medicinal plant in tropical countries. Through of Dilution Testing in Multiple Tubes System, it were determined the intensity of bacterial inhibition activity (IINIB/ Bacteriostasy) and the intensity of bacterial inactivation activity (IINAB/ Bactericidie) of alcoholic extracts of two accesses of hibiscus, known as: Palmares do Sul/RS and Porto Alegre/RS on standardized inoculum of Enterococcus faecalis (ATCC 19433), Escherichia coli (ATCC 11229), Salmonella Enteritidis (ATCC 11076) and Staphylococcus aureus (ATCC 25923). At the same time, the total content of polyphenols and anthocyanins in the calyxes and fruits with seeds hibiscus was determined. The antimicrobial activity of alcoholic extract of the calyxes in both accesses showed a significant positive difference when related to the alcoholic extract of the fruits with seeds. Salmonella enteritidis was the most sensitive bacteria to the alcoholic extract of calyxes of the hibiscus while the most resistant was Staphylococcus aureus, independent of the variable access, considering only the plant structure. In relation to the alcoholic extract of the fruits with seeds, Escherichia coli showed the highest sensitivity and Staphylococcus aureus the highest resistence. Total polyphenols and anthocyanins of alcoholic extract of calyxes‟s values showed a significant difference and they were superior to alcohol extract of fruits with seeds. Possibly there is a direct relationship between the concentration of anthocyanin and antibacterial activity in different structures of the hibiscus plant.

O Yacon (Smallanthus sonchifolius) é uma planta que pertence à família Asteraceae, é originário das montanhas dos Andes e no Brasil seu cultivo iniciou-se em 1991. Possui carboidratos solúveis tais como frutose, glicose, sacarose e frutooligossacarídeos (FOS), sendo que os FOS não podem ser metabolizados pelo trato digestivo humano, tendo dessa forma atividade prebiótica. Este trabalho visou estudar o efeito do branqueamento no yacon e posterior secagem mediante ar quente. As raízes foram limpas e selecionadas considerando a ausência de injúrias visuais e infecções. A seguir foram descascadas e cortadas em forma de rodelas (espessuras de 1,75 ± 0,35 mm) e cubos (1,00 ± 0,01cm3). Foi verificada a ocorrência da solubilização dos açúcares durante o branqueamento, onde foram avaliadas as perdas de inulina, glicose e frutose em diferentes condições de tempo e temperatura. Foi observada a maior solubilização nas amostras em rodelas que em cubos na maioria dos tratamentos estudados. O teste de Tukey indicou que no branqueamento do yacon em forma de rodelas e cubos, o tempo, a temperatura e a interação entre eles foi significativa na solubilização dos açúcares, exceto na frutose (nas amostras em rodelas) e na inulina (nas amostras em cubos) onde somente foi significativo o tempo e a temperatura. Os resultados obtidos da superfície de resposta permitiram obter modelos estatísticos para estimar a perda de açúcares no branqueamento das amostras em rodelas, estimando as condições de maior solubilização. Devido a essas perdas, estudou-se o branqueamento a vapor em amostras em rodelas, que foram colocadas dentro de uma autoclave gerando vapor a 100oC nos tempos de 1, 2, 4, 6, 8 e 10 minutos, sendo a melhor condição a 4 minutos, onde foi possível reduzir a atividade enzimática da PER e PPO em 84,6% e 83,7%, correspondendo a perdas de inulina, glicose e frutose de 30,6, 39,4 e 15,8% respectivamente. A seguir foi realizada a secagem nas amostras de yacon, nas temperaturas de 50, 60 e 70°C por 5 horas e 30 minutos sem e com branqueamento, onde verificou-se o efeito do pré-tratamento e da temperatura sobre a redução da umidade e da atividade de água, revelando que o menor tempo de secagem foi obtido a 70°C em amostras com branqueamento. Também foi observado que após 5 horas de secagem a concentração de inulina diminuiu, enquanto que as concentrações de glicose e frutose aumentaram, sendo que os teores desses componentes no final da secagem não diferiram com a temperatura, tanto nas amostras que sofreram ou não branqueamento. No entanto, houve conversão dos FOS em açúcares redutores. O aumento na concentração dos açúcares redutores pode ser devido à presença de atividade enzimática da inulinase. / Yacon (Smallanthus sonchifolius) is a plant belonging to the Asteraceae family and originated in the Andes Mountains, having been cultivated in Brazil since 1991. It contains soluble carbohydrates such as fructose, glucose, sucrose and fructooligosaccharides (FOS), the latter not being metabolized in the human digestive tract and thus presenting prebiotic activity. This work aimed to study the effect of blanching and subsequent hot air drying on yacon. The roots were cleaned and selected considering the absence of visual injury and infections. They were then peeled and cut into slices (1.75 ± 0.35 mm thick) and cubes (1.00 ± 0.01cm2). The sugars were shown to dissolve during blanching, and the losses of inulin, glucose and fructose were determined under different conditions of time and temperature. For the majority of conditions studied, greater dissolution was observed with the slices than with the cubes. Tukey's test indicated that both the time and the temperature and the interaction between them were significant with respect to the dissolution of sugars in the blanching of yacon in the form of both slices and cubes, with the exception of fructose (for the sliced samples) and inulin (for the samples in cubes), where only the time and temperature were significant. The results obtained from the response surface allowed for the production of statistical models to estimate the loss of sugars during blanching for the samples in slices, estimating the conditions for greatest dissolution. Due to these losses, steam blanching of the slices was studied, placing the slices inside an autoclave generating steam at 100ºC for times of 1, 2, 4, 6, 8 and 10 minutes, the best condition being that of 4 minutes where it was possible to reduce the PER and PPO activities by 84.6% and 83.7%, respectively, with losses of inulin, glucose and fructose of 30.6, 39.4 and 15.8%, respectively. Drying of the yacon samples at temperatures of 50, 60 and 70ºC for 5 hours and 30 minutes, with and without blanching, was then carried out, verifying the effect of the pre-treatment and of the drying temperature on the reduction in moisture content and water activity. The shortest drying time was obtained at 70ºC with blanched samples. It was also observed that after 5 hours of drying the concentration of inulin decreased, whereas the concetrations of glucose and fructose increased, the contents of these components at the end of the drying period not varying according to the drying temperature, for either the blanched or non-blanched samples. Thus the FOS were converted into reducing sugars, and the increase in reducing sugars could have been due to the presence of inulinase activity.

Através de Testes de Diluição em Sistema de Tubos Múltiplos determinou-se a Intensidade de Atividade de Inibição Bacteriana (IINIB/bacteriostasia) e a Intensidade de Atividade de Inativação Bacteriana (IINAB/bactericidia) de soluções contendo extratos hidroetanólicos e hídricos (decocto e infuso) de três acessos de Bixa orellana L. (urucum) a saber: Arroio do Meio/RS, Eldorado do Sul/RS e Maringá/PR, sobre inóculos padronizados de Salmonella Enteritidis (ATCC 11076), Escherichia coli (ATCC 11229), Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 19433) e Listeria monocytogenes (ATCC 19114). Determinou-se, paralelamente, o teor de bixina presente nas sementes. Os extratos hídricos apresentaram baixa atividade de inibição e/ou inativação sobre os inóculos bacterianos, enquanto que a forma de extração hidroetanólica apresentou atividade antibacteriana seletiva e significativamente mais intensa (inibição/inativação) entre as cinco bactérias testadas. Independente da forma de extração, as bactérias Enterococcus faecalis e Listeria monocytogenes foram as mais sensíveis à atividade antibacteriana, enquanto que Escherichia coli apresentou a menor sensibilidade. Houve diferença significativa entre os teores de bixina dos três acessos, e, consequentemente, a atividade antibacteriana determinada mostrou-se diretamente proporcional a estes teores. / Through of Dilution Tests in Multiple Tubes System it was determined the intensity of bacterial inhibition activity (IINIB/bacteriostasy) and the intensity of bacterial inactivation activity (IINAB/bactericidie) from solutions containing hidroetanolic and hydric (decoction and infusion) extracts of tree accesses from Bixa orellana L. (annatto) at know: Arroio do Meio/RS, Eldorado do Sul/RS and Maringá/PR, on standardized inocula of Salmonella Enteritidis (ATCC 11076), Escherichia coli (ATCC 11229), Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 19433) and Listeria monocytogenes (ATCC 19114). It was determined, in parallel, the content of bixin present in the three different accesses of the seeds. The forms of hydric extraction showed low inhibition and/or inactivation activity of the bacterial inocula, and the hidroetanolic extract form showed selective antibacterial activity and significantly pronounced inhibition/inactivation against the five bacteria tested. Independent of the extraction forms, Enterococcus faecalis and Listeria monocytogenes were the more sensitive agents to the antibacterial activity. Escherichia coli had the lowest sensitivity to all forms of extraction. The bixin contents were significantly different between the accesses and, consequently, the antibacterial activity was directly proportional to this contents.