Orientador: Valdir Augusto Neves / Banca: Edwil Aparecida de Lucca Gattás / Banca: Eduardo Purgatto / Resumo: Os frutos e vegetais apresentam-se fisiologicamente ativos após a colheita, dessa forma as perdas pós-colheita ocorrem pela influência de diferentes fatores onde se destacam a respiração, a temperatura, a umidade, a concentração de oxigênio e gás carbônico, a produção de etileno e a ação de enzimas endógenas associadas com processos de deterioração. As raízes de yacon têm sido cada vez mais consumidas devido à revelação de qualidades medicinais. A manutenção de sua qualidade in natura é um sério problema nos processos pós-colheita devido a inúmeras reações metabólicas. A peroxidase (POD, E.C.1.11.1.7) é largamente encontrada nos vegetais apresentando importante papel fisiológico/bioquímico embora uma precisa função não tenha ainda sido estabelecida. Sua importância para ciência dos alimentos evidencia-se pelas relações com alterações indesejáveis na qualidade e resistência dos vegetais, tornando interessante sua supressão parcial ou total no pós-colheita. Os objetivos desse trabalho foram isolar, purificar e caracterizar a peroxidase de raízes de yacon. Condições de extração para a POD de yacon foram estabelecidas e a enzima foi isolada por precipitação com sulfato de amônio, eluição em Sephadex G-25 e DEAE-celulose. Somente um pico de atividade foi eluído no processo de purificação com um fator de purificação de 222,33. O peso molecular determinado foi 34.8 kDa com valores de pH e temperatura ótima de 5,5 e 35°C, respectivamente. As constantes Km e Vmax foram de 14,227 mM e 17409 UA/mL, para s-dianisidina, e de 14,434 mM e 14830 UA/mL, para H2O2. Foram testados os efeitos inibidores de sais, quelantes, compostos sulfidrila, ácidos fenólicos e outros. Estudos de inativação térmica da POD foram realizados, nos quais se verificou o efeito protetor da sacarose na inativação enzimática e o efeito promotor de inativação pela ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The fruits and vegetables still continue physiologically active after the crop, in that way the postharvest losses occurs by the influence of different factors, like the respiration, temperature, humidity, concentration of oxygen and carbonic gas, ethylene production and the action of endogenous enzymes associated with deterioration processes. Yacon (Smallanthus sonchifolius) roots has been more and more consumed by the revelation of prebiotic and medicinal properties. The maintenance of its quality in the postharvest is a serious problem due to several metabolic reactions. Peroxidase (POD, E.C.1.11.1.7) is widely found in plants, having physiological/biochemical importance, although a precise function has not still been established. The POD importance for food science is due to its relationships with alteration of the quality and resistance of the vegetables to the postharvest factors. Its total or partial suppression by thermal inactivation or inhibitor compounds may be interesting for postharvest interests. The objectives of this work were to isolate, purify and characterize the POD from yacon roots. Extraction conditions for yacon POD were determined and the enzyme was isolated by precipitation with ammonium sulfate, elution on Sephadex G-25 and DEAE-cellulose. Only one peak of enzyme activity was eluted on the purification process with a 222.33 fold purification factor. The determined molecular weight was 34.8 kDa. The optimum pH and temperature values were pH 5.5 and 35°C, respectively. Km and Vmax for ï-dianisidine was 14.227 mM and 17409 UA/mL, and 14.434 mM and 14830 UA/mL for H2O2, respectively. The effects of metals, chelating agents, sulphidryl, phenolic acids and other compounds as POD inhibitors were evaluated... (Complete abstract, click electronic address below). / Mestre

Made available in DSpace on 2014-06-11T19:23:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-06-02Bitstream added on 2014-06-13T19:29:44Z : No. of bitstreams: 1 kamimura_gkf_me_arafcf.pdf: 713918 bytes, checksum: 90d6ccb478cdaac799bbb870832f99ec (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Os frutos e vegetais apresentam-se fisiologicamente ativos após a colheita, dessa forma as perdas pós-colheita ocorrem pela influência de diferentes fatores onde se destacam a respiração, a temperatura, a umidade, a concentração de oxigênio e gás carbônico, a produção de etileno e a ação de enzimas endógenas associadas com processos de deterioração. As raízes de yacon têm sido cada vez mais consumidas devido à revelação de qualidades medicinais. A manutenção de sua qualidade in natura é um sério problema nos processos pós-colheita devido a inúmeras reações metabólicas. A peroxidase (POD, E.C.1.11.1.7) é largamente encontrada nos vegetais apresentando importante papel fisiológico/bioquímico embora uma precisa função não tenha ainda sido estabelecida. Sua importância para ciência dos alimentos evidencia-se pelas relações com alterações indesejáveis na qualidade e resistência dos vegetais, tornando interessante sua supressão parcial ou total no pós-colheita. Os objetivos desse trabalho foram isolar, purificar e caracterizar a peroxidase de raízes de yacon. Condições de extração para a POD de yacon foram estabelecidas e a enzima foi isolada por precipitação com sulfato de amônio, eluição em Sephadex G-25 e DEAE-celulose. Somente um pico de atividade foi eluído no processo de purificação com um fator de purificação de 222,33. O peso molecular determinado foi 34.8 kDa com valores de pH e temperatura ótima de 5,5 e 35°C, respectivamente. As constantes Km e Vmax foram de 14,227 mM e 17409 UA/mL, para s-dianisidina, e de 14,434 mM e 14830 UA/mL, para H2O2. Foram testados os efeitos inibidores de sais, quelantes, compostos sulfidrila, ácidos fenólicos e outros. Estudos de inativação térmica da POD foram realizados, nos quais se verificou o efeito protetor da sacarose na inativação enzimática e o efeito promotor de inativação pela... / The fruits and vegetables still continue physiologically active after the crop, in that way the postharvest losses occurs by the influence of different factors, like the respiration, temperature, humidity, concentration of oxygen and carbonic gas, ethylene production and the action of endogenous enzymes associated with deterioration processes. Yacon (Smallanthus sonchifolius) roots has been more and more consumed by the revelation of prebiotic and medicinal properties. The maintenance of its quality in the postharvest is a serious problem due to several metabolic reactions. Peroxidase (POD, E.C.1.11.1.7) is widely found in plants, having physiological/biochemical importance, although a precise function has not still been established. The POD importance for food science is due to its relationships with alteration of the quality and resistance of the vegetables to the postharvest factors. Its total or partial suppression by thermal inactivation or inhibitor compounds may be interesting for postharvest interests. The objectives of this work were to isolate, purify and characterize the POD from yacon roots. Extraction conditions for yacon POD were determined and the enzyme was isolated by precipitation with ammonium sulfate, elution on Sephadex G-25 and DEAE-cellulose. Only one peak of enzyme activity was eluted on the purification process with a 222.33 fold purification factor. The determined molecular weight was 34.8 kDa. The optimum pH and temperature values were pH 5.5 and 35°C, respectively. Km and Vmax for ï-dianisidine was 14.227 mM and 17409 UA/mL, and 14.434 mM and 14830 UA/mL for H2O2, respectively. The effects of metals, chelating agents, sulphidryl, phenolic acids and other compounds as POD inhibitors were evaluated... (Complete abstract, click electronic address below).

An array of Bacillus licheniformis strains were isolated from a commercial powdered milk process. Bacteriophages exhibiting activity against B. licheniformis were isolated from cattle manure and effluent samples destined for a lagoon at a dairy farm. After sequencing, 8 of the 10 phages were found to be novel and genetically differentiated. Transmission electron scanning microscopy (TSEM) was performed. All bacteriophages were of the family Herelleviridae with contractile tail sheaths ranging from 80µm to 150µm and, surprisingly, survived a common fluid milk processing treatment used to inactivate vegetative cells. The survival of the phage after high temperature short time pasteurization of 73℃ for 20 s shows that the use of bacteriophages in milk to control B. licheniformis could be applied as a potential quality control, retarding the germination of spores and reduction of final spore counts in products with long run times such as dairy powders.

Considerando a necessidade da pasteurização da água de coco verde, após a abertura do fruto, devido à ação de microrganismos e das enzimas presentes, peroxidase (POD) e polifenoloxidase (PFD), o presente trabalho tem como objetivo estudar o comportamento da atividade dessas enzimas, através de processo térmico contínuo. Na primeira parte foi estudada a cinética de inativação térmica das enzimas em processo de pasteurização da água de coco, realizado em trocador de calor a placas (TCP) de laboratório, ARMFIELD FT 43–A, a temperatura de 90°C em diferentes tempos de retenção. Amostras da água de coco não processada e processada foram congeladas a – 30ºC para as análises enzimáticas. Na segunda parte, com base nos resultados obtidos de cinética, foram realizados processos a 90 ºC com tempos de retenção de 120, 180 e 200 s para o estudo do comportamento dessas enzimas durante o armazenamento. Análises de pH, acidez, turbidez e atividade enzimática também foram realizadas durante o tempo de armazenamento de amostras da água de coco processada e não processada em temperatura ambiente, refrigerada (10ºC) e congelada (- 20ºC). Cada lote de água de coco foi caracterizado pela densidade, teores de sólidos totais e solúveis, cinzas, pH, acidez e turbidez. Nos processos térmicos contínuos realizados a 90ºC a POD e PFD presentes na água de coco apresentaram pelo menos duas isoenzimas com estabilidades diferentes e a PFD mostrou-se mais termorresistente que a POD. A POD presente na água de coco processada, durante o armazenamento a – 30ºC, apresentou-se mais estável que a PFD. A água de coco processada que apresentou maior estabilidade físico-química e enzimática foi a armazenada congelada. / Considering the need for green coconut water pasteurization after opening the fruit due to microbial and enzymatic (peroxidase (POD) and polyphenol oxidase (PFD)) action, the objective of this work is to study the behavior of the activity of these enzymes during continuous thermal processing. The first stage of the work was the study of the enzymes thermal inactivation kinetics during continuous pasteurization of coconut water, performed in a laboratory plate heat exchanger, Armfield 43 – A, at a temperature of 90°C with different holding times. Samples of unprocessed and processed coconut water were frozen at - 30°C for enzymatic analyses. In the second stage, based on the results obtained, processing was performed at 90°C with holding times of 80, 120, 180 and 200 s for studying the behavior of these enzymes during storage. Analyses of the pH, titratable acidity, turbidity and enzymatic activity were conducted during storage in processed and unprocessed samples kept at ambient temperature, refrigerated (10°C) and frozen (- 20°C). Each coconut water batch was characterized by density, total and soluble solids, ashes, pH, titratable acidity and turbidity. For the continuous thermal processes carried out at 90°C both POD and PFD presented at least two isozymes with different thermal stability, and PFD was more thermally resistant than POD. During storage at - 30°C POD in processed coconut water was more stable than PFD. The highest physicochemical and enzymatic stability was achieved for processed coconut water under frozen storage.

A pasteurização do leite destinado ao consumo é obrigatória no Brasil e o sistema rápido (75ºC/15 a 20 segundos) é o mais empregado no país. O processo visa eliminar. Os parâmetros de tempo e temperatura empregados no mundo foram definidos após estudos sobre a resistência térmica do Mycobacterium tuberculosis e da Coxiella burnetti, reconhecidos como os microrganismos patogênicos, não formadores de esporos e que eventualmente podem estar presentes no leite cru, que apresentam a maior resistência térmica. Entretanto, não há estudos sobre a resistência térmica do M. bovis que circula nos bovinos no Brasil. Este estudo propôs-se a avaliar a resistência térmica (75ºC) de cinco espoligotipos de M. bovis, isolados de bovinos abatidos no estado de São Paulo, em leite integral experimentalmente contaminado. Leite UHT foi contaminado com M. bovis e, então, submetido a tratamento térmico em banho-maria a 75ºC por 20 segundos. Cada espoligotipo foi testado 3 vezes. As amostras foram retiradas do banho nos tempos 0 (o momento em que o leite atingiu 75ºC), 5, 10, 15 e 20, correspondendo ao tempo, em segundos, de tratamento térmico. O leite contaminado também foi analisado, para quantificação da carga inicial. O controle do processo envolveu o acompanhamento da temperatura do leite (um tubo com termômetro) e análise das enzimas fosfatase alcalina e peroxidase ao final do tratamento; para tal, amostras de leite cru foram tratadas juntamente com as amostras-teste. Para quantificação, foi realizada a diluição decimal seriada seguida da semeadura em duplicata em meio Stonebrink-Leslie (37ºC/45dias). Os resultados mostraram que foi na fase de aquecimento que ocorreu a maior taxa de morte de todos os espoligotipos. Houve diferença de resistência entre os espoligotipos ao processo que simulou a pasteurização rápida e o espoligotipo BR024 foi o mais resistente. Conclui-se que houve diferença da eficácia da pasteurização, de acordo com o espoligotipo testado, mas que os resultados precisam ser investigados mais detalhadamente. / The pasteurization of milk for consumption is mandatory in Brazil and fast system (75 ° C/15 to 20 seconds) is the most used around the country. The process aims to eliminate. The parameters of time and temperature were set in the world after studies on the thermal resistance of Mycobacterium tuberculosis and Coxiella burnetii, recognized as the pathogenic microorganisms, not spore-forming and eventually may be present in raw milk, with the strongest resistance heat. However, no studies on the thermal resistance of M. bovis circulating in cattle in Brazil. This study aimed to evaluate the thermal resistance (75º C) of five espoligotipos M. bovis isolated from cattle slaughtered in the state of Sao Paulo in experimentally infected whole milk. UHT milk was contaminated with M. bovis and then subjected to heat treatment in a water bath at 75 º C for 20 seconds. Each espoligotipo was tested 3 times. Samples were taken from the bath at 0 (the time when the milk reached 75 ° C), 5, 10, 15 and 20, corresponding to time, in seconds, the heat treatment. The contaminated milk was also analyzed to quantify the initial charge. The control process involves monitoring the temperature of the milk (a tube with a thermometer) and analysis of the enzymes alkaline phosphates and peroxidase the end of treatment, for such raw milk samples were treated with the test samples. For quantification, we performed a ten-fold dilution serial followed by seeding in duplicate in the middle Stonebrink-Leslie (37 C/45dias). The results showed that it was warming up that had the highest death rate of all espoligotipos. There were differences in resistance between espoligotipos the process that simulated pasteurization and rapid espoligotipo br024 was the toughest. It was concluded that there was no difference of the effectiveness of pasteurization, according to the espoligotipo tested, but the results need to be investigated further.

A pasteurização do leite destinado ao consumo é obrigatória no Brasil e o sistema rápido (75ºC/15 a 20 segundos) é o mais empregado no país. O processo visa eliminar. Os parâmetros de tempo e temperatura empregados no mundo foram definidos após estudos sobre a resistência térmica do Mycobacterium tuberculosis e da Coxiella burnetti, reconhecidos como os microrganismos patogênicos, não formadores de esporos e que eventualmente podem estar presentes no leite cru, que apresentam a maior resistência térmica. Entretanto, não há estudos sobre a resistência térmica do M. bovis que circula nos bovinos no Brasil. Este estudo propôs-se a avaliar a resistência térmica (75ºC) de cinco espoligotipos de M. bovis, isolados de bovinos abatidos no estado de São Paulo, em leite integral experimentalmente contaminado. Leite UHT foi contaminado com M. bovis e, então, submetido a tratamento térmico em banho-maria a 75ºC por 20 segundos. Cada espoligotipo foi testado 3 vezes. As amostras foram retiradas do banho nos tempos 0 (o momento em que o leite atingiu 75ºC), 5, 10, 15 e 20, correspondendo ao tempo, em segundos, de tratamento térmico. O leite contaminado também foi analisado, para quantificação da carga inicial. O controle do processo envolveu o acompanhamento da temperatura do leite (um tubo com termômetro) e análise das enzimas fosfatase alcalina e peroxidase ao final do tratamento; para tal, amostras de leite cru foram tratadas juntamente com as amostras-teste. Para quantificação, foi realizada a diluição decimal seriada seguida da semeadura em duplicata em meio Stonebrink-Leslie (37ºC/45dias). Os resultados mostraram que foi na fase de aquecimento que ocorreu a maior taxa de morte de todos os espoligotipos. Houve diferença de resistência entre os espoligotipos ao processo que simulou a pasteurização rápida e o espoligotipo BR024 foi o mais resistente. Conclui-se que houve diferença da eficácia da pasteurização, de acordo com o espoligotipo testado, mas que os resultados precisam ser investigados mais detalhadamente. / The pasteurization of milk for consumption is mandatory in Brazil and fast system (75 ° C/15 to 20 seconds) is the most used around the country. The process aims to eliminate. The parameters of time and temperature were set in the world after studies on the thermal resistance of Mycobacterium tuberculosis and Coxiella burnetii, recognized as the pathogenic microorganisms, not spore-forming and eventually may be present in raw milk, with the strongest resistance heat. However, no studies on the thermal resistance of M. bovis circulating in cattle in Brazil. This study aimed to evaluate the thermal resistance (75º C) of five espoligotipos M. bovis isolated from cattle slaughtered in the state of Sao Paulo in experimentally infected whole milk. UHT milk was contaminated with M. bovis and then subjected to heat treatment in a water bath at 75 º C for 20 seconds. Each espoligotipo was tested 3 times. Samples were taken from the bath at 0 (the time when the milk reached 75 ° C), 5, 10, 15 and 20, corresponding to time, in seconds, the heat treatment. The contaminated milk was also analyzed to quantify the initial charge. The control process involves monitoring the temperature of the milk (a tube with a thermometer) and analysis of the enzymes alkaline phosphates and peroxidase the end of treatment, for such raw milk samples were treated with the test samples. For quantification, we performed a ten-fold dilution serial followed by seeding in duplicate in the middle Stonebrink-Leslie (37 C/45dias). The results showed that it was warming up that had the highest death rate of all espoligotipos. There were differences in resistance between espoligotipos the process that simulated pasteurization and rapid espoligotipo br024 was the toughest. It was concluded that there was no difference of the effectiveness of pasteurization, according to the espoligotipo tested, but the results need to be investigated further.

High-pressure needleless injection (HPNI) is a process where small-diameter, high-velocity burst of liquid, penetrate foods at pressures ≤ 10,000 psi. The potential of HPNI as an enhancing technique for meat was studied. In study 1, HPNI translocated surface E. coli O157 into the interior of beef eye-of-round subprimals with an incidence of 40 (±7), 25 (±8), and 25 (±8)% for meat that had been surface-inoculated with a four-strain cocktail at 0.5, 1, and 2 log10 CFU/cm2, respectively. Run-off water contained 2, 2, and 3 log10 CFU/ml and was used for HPNI of additional subprimals, which resulted in a cross-contamination incidence of 83 (±4), 60 (±15), and 37 (±6) %, respectively. Incidence of translocation and cross-contamination was similar at all sampled levels below the inoculated surface. Study 1 results indicate that surface microflora will be translocated from the surface into the interior of HPNI-treated beef by the injection fluid and by cross-contamination with recycled fluid. In study 2, E. coli was undetected in cooked steaks (63˚C internal) cut from subprimals inoculated with 2 log10 CFU/cm2 and HPNI processed (study 1). Although cooking reduced E. coli counts, determination of complete kill was not possible because the detection limit for bacterial recovery was about 1 log10 CFU/g. Steaks cut from HPNI-processed subprimals took longer (p <0.05) to reach 63˚C with grilling or broiling, compared to control steaks, possibly due to increased moisture in enhanced steaks. In study 3, sensory acceptance of steaks was evaluated by a consumer panel. Appearance, flavor, and overall acceptance were similar among the untreated control, HPNI steaks, blade tenderized steaks (BT steaks), and steaks cut from subprimals that had been needle-injected with 0.35% (wt/vol) sodium tripolyphosphate using needle injection (NI-subprimal steaks) or HPNI (HPNI-subprimal steaks). Texture of BT steaks (6.5±1.9) was more liked than control steaks (5.8±1.8), while texture was similar for all other comparisons. Conversely, Warner-Bratzler shear force was NI-subprimal steaks < control < HPNI steaks = HPNI-subprimal steaks = BT steaks. Lack of correspondence between texture acceptance data and WBSF suggests that sensory scores were influenced by factors other than the force required for mechanical shear.

Considerando a necessidade da pasteurização da água de coco verde, após a abertura do fruto, devido à ação de microrganismos e das enzimas presentes, peroxidase (POD) e polifenoloxidase (PFD), o presente trabalho tem como objetivo estudar o comportamento da atividade dessas enzimas, através de processo térmico contínuo. Na primeira parte foi estudada a cinética de inativação térmica das enzimas em processo de pasteurização da água de coco, realizado em trocador de calor a placas (TCP) de laboratório, ARMFIELD FT 43–A, a temperatura de 90°C em diferentes tempos de retenção. Amostras da água de coco não processada e processada foram congeladas a – 30ºC para as análises enzimáticas. Na segunda parte, com base nos resultados obtidos de cinética, foram realizados processos a 90 ºC com tempos de retenção de 120, 180 e 200 s para o estudo do comportamento dessas enzimas durante o armazenamento. Análises de pH, acidez, turbidez e atividade enzimática também foram realizadas durante o tempo de armazenamento de amostras da água de coco processada e não processada em temperatura ambiente, refrigerada (10ºC) e congelada (- 20ºC). Cada lote de água de coco foi caracterizado pela densidade, teores de sólidos totais e solúveis, cinzas, pH, acidez e turbidez. Nos processos térmicos contínuos realizados a 90ºC a POD e PFD presentes na água de coco apresentaram pelo menos duas isoenzimas com estabilidades diferentes e a PFD mostrou-se mais termorresistente que a POD. A POD presente na água de coco processada, durante o armazenamento a – 30ºC, apresentou-se mais estável que a PFD. A água de coco processada que apresentou maior estabilidade físico-química e enzimática foi a armazenada congelada. / Considering the need for green coconut water pasteurization after opening the fruit due to microbial and enzymatic (peroxidase (POD) and polyphenol oxidase (PFD)) action, the objective of this work is to study the behavior of the activity of these enzymes during continuous thermal processing. The first stage of the work was the study of the enzymes thermal inactivation kinetics during continuous pasteurization of coconut water, performed in a laboratory plate heat exchanger, Armfield 43 – A, at a temperature of 90°C with different holding times. Samples of unprocessed and processed coconut water were frozen at - 30°C for enzymatic analyses. In the second stage, based on the results obtained, processing was performed at 90°C with holding times of 80, 120, 180 and 200 s for studying the behavior of these enzymes during storage. Analyses of the pH, titratable acidity, turbidity and enzymatic activity were conducted during storage in processed and unprocessed samples kept at ambient temperature, refrigerated (10°C) and frozen (- 20°C). Each coconut water batch was characterized by density, total and soluble solids, ashes, pH, titratable acidity and turbidity. For the continuous thermal processes carried out at 90°C both POD and PFD presented at least two isozymes with different thermal stability, and PFD was more thermally resistant than POD. During storage at - 30°C POD in processed coconut water was more stable than PFD. The highest physicochemical and enzymatic stability was achieved for processed coconut water under frozen storage.

No description available.

Contaminated wheat flour has been identified as the probable vehicle of a multi-state outbreak of Escherichia coli O157:H7 associated with ready-to-bake cookie dough. Several cookie dough manufacturers are currently using heat-treated flour for ready-to-bake products, although data on thermal inactivation of foodborne pathogens in wheat flour remains scarce. The objective of this research was to first determine appropriate methods and parameters for bacterial inoculation and thermal treatment of wheat flour, and to subsequently determine the population reductions of E. coli O157:H7 and Salmonella Agona in artificially contaminated wheat flour following thermal treatment for 1, 5, 15 or 30 minutes at 55, 60, 65 or 70°C in a shaking water bath. Flour samples (aw = 0.55) in sterile plastic bags were individually inoculated (~109 CFU/g), pulsified to distribute cultures, and pressed to a uniform thickness (1mm) prior to heat treatment. Following treatment, samples were rapidly cooled and diluted with peptone water; then plated onto Tryptic Soy Agar (TSA) and incubated at 37°C for 24 h prior to enumeration. The minimum heat treatments required for a 5-log reduction in microbial populations (~ 109CFU/g to ~ 104CFU/g) were 5 minutes at 70°C and 30 minutes at 70°C for E. coli O157:H7 and S. Agona, respectively. This research supports the hypothesis that the microbiological safety of ready-to-bake products may be improved by the use of heat-treated flour. / Master of Science in Life Sciences