Heat treatment (around 50&deg / C) is a major step in honey filling and packaging that is applied before filtration to decrease viscosity, reduce the moisture level, to destroy yeasts, liquefy crystals and delay crystallization. As a result, formation of Hydroxy Methyl Furfural (HMF), decrease in enzymatic activity, color deterioration, decrease in viscosity and many other structural changes are observed. HMF is produced as a result of Maillard reaction and/or hexose dehydration -which is undesirable-, practically, it is found in fresh honey in low levels, and increases due to heat treatment, storage temperature, pH (acidity) and sugar concentration of honey. HMF level and diastase number are important quality parameters and shelf life indicators of honey. Alternatives of v heat treatment may be the use of ultrasound and high hydrostatic pressure (HHP) to decrease viscosity, liquefy honey and thus minimise adverse affects of heat treatment. Therefore, the aim of this study is to evaluate the effect of HHP (220-330 MPa, 50-60&deg / C, time) and ultrasound (24 kHz) on liquefaction and quality parameters (HMF, diastase number, color and viscosity) of different honey varieties (sunflower, cotton and canola) and to compare the changes with heat treated (50&deg / C and 60&deg / C, time) and untreated honey. Based on the results of the chemical and physical analysis, for HHP treatment the best treatment combination was determined as 220 MPa, 50&deg / C, 106 min. For ultrasound treatment the best treatment combinations were determined as 7 mm probe- 0.5 cycle (batch) applications. On this basis the study points out that Ultrasound and HHP can be suggested as alternative methods to traditional thermal treatment for the liquefaction of honey crystals. When compared to thermal treatment, Ultrasound is advantageous in shorter application times, slight changes in quality parameters and ease in operation. HHP treatment is also an alternative method with shorter application times and lower HMF values.

High Hydrostatic Pressure (HHP) application, alone or in combination with refrigeration, ambient or moderate heating temperatures / inactivates pathogenic and spoilage microorganisms and conserves the product &bdquo / &bdquo / freshness

Orientadores: Carlos Francisco Sampaio Bonafé, Cláudio Chrysostomo Werneck / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-17T12:27:33Z (GMT). No. of bitstreams: 1 Carvalho_RicardoDuraesde_M.pdf: 7651307 bytes, checksum: 3194190cf6a33de3b0cdc4ddb7424afd (MD5) Previous issue date: 2010 / Resumo: A exposição à alta pressão é um método eficiente na inativação bacteriana, o qual é importante para a esterilização de alimentos. Neste trabalho, examinamos a inativação por alta pressão hidrostática em Aeromonas hydrophila AH 191, produtora de aerolisina, a qual possui atividades citotóxica, enterotóxica e hemolítica. Trinta min a 250 MPa (25 °C em PBS 0,1 M, pH 7,4), resultou em 9 log de UFC/mL de inativação bacteriana (a partir de 109 UFC/mL para <10 UFC/mL), porém, os tratamentos não alteraram a atividade hemolítica, enterotóxica e citotóxica produzidas por A. hydrophila AH 191. O sobrenadante de cultura bacteriana e o leite contaminado por A. hydrophila AH 191, pressurizado ou não, foram aplicados sobre células Vero e CaCo2, suspensões de hemácias humanas e de carneiro, e injetadas por via oral gástrica em camundongos Swiss, reproduzindo o ensaio de Dean. Os resultados mostram que os títulos das toxinas em amostras pressurizadas permaneceram iguais aos do controle, assim como as toxinas presentes nos sobrenadantes de cultura e no leite foram capazes de produzir acúmulo de líquidos no intestino de camundongos recém-nascidos. Estes resultados indicam a relevância do estudo na inativação de bactérias para a segurança alimentar e realça a importância de cautela no uso de alta pressão para a esterilização de alimentos, pois embora as bactérias possam ser inativadas, as toxinas produzidas por este micro-organismos podem continuar ativas, representando uma ameaça à saúde humana / Abstract: Exposure to high pressure is an efficient method for bacterial inactivation, which is important for the food sterilization. We examine the inactivation by high hydrostatic pressure in A. hydrophila AH 191, producer of aerolysin, which has cytotoxic, enterotoxic and hemolytic activities. A 30 min treatment at 250 MPa (25 °C in 0.1 M PBS, pH 7,4) resulted in 9 log CFU/mL of bacteria inactivation (from 109 CFU/mL to <10 CFU/mL). However, the treatment did not inactivate the hemolytic, enterotoxic and cytotoxic activities produced by A. hydrophila AH 191. The supernatant of bacterial culture and milk contaminated with A. hydrophila AH 191 pressurized or not were applied to Vero and CaCo2 cells, suspensions of human and sheep erythrocytes, and injected into mice via oral gastric Swiss reproducing the Dean test. The results show that evidence of toxins in samples pressurized remained similar to that of control, and the toxins in culture supernatants and milk were able to produce fluid accumulation in the intestine of suckling mouse. These results indicate the relevance of the study for bacteria inactivation to food security and enhance the importance for caution in the use of high prssure in foods sterilization, since although the bacteria can be inactivated, the toxins produced by micro-organisms continue to be active, representing a human health threat / Mestrado / Bioquimica / Mestre em Biologia Funcional e Molecular

Orientador: Carlos Francisco Sampaio Bonafe / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-11T01:21:44Z (GMT). No. of bitstreams: 1 Santos_JoseLuisdaRocha_D.pdf: 2961900 bytes, checksum: 8d0413331d106601d34efccd4f8e6bb7 (MD5) Previous issue date: 2008 / Resumo: Os vírus são estruturas auto-associaveis muito eficientes, porém, pouco é conhecido sobre a termodinâmica que governa essa associação dirigida. Em altos níveis de pressão ocorre dissociação do TMV ou desnaturação quando a pressão é combinada com uréia. Para um melhor entendimento de tais processos nós calculamos os parâmetros termodinâmicos aparentes de dissociação e desnaturação do TMV, assumindo uma condição de estado estacionário "steady-state condition". Esses processos podem ser monitorados pela diminuição de espalhamento de luz da solução virai devido ao processo de dissociação e pelo desvio para o vermelho do espectro de emissão de fluorescência, que ocorre com o processo de desnaturação. Nós determinamos a estequiometria aparente de uréia considerando a reação de equilíbrio de dissociação e desnaturação das subunidades do TMV, as quais forneceram, respectivamente, 1,53 e 11,1 mols de uréia/mol de subunidade de TMV. As condições de dissociação e desnaturação foram atingidas em uma via próxima da reversível, permitindo a determinação de parâmetros termodinâmicos. Analises de gel filtração em HPLC, microscopia eletrônica e dicroísmo circular confirmaram os processos de dissociação e desnaturação. Os cálculos das estequiometrias aparentes de uréia de vários vírus baseados em resultados espectroscópicos de artigos recentes mostraram que os processos de dissociação e desnaturação seguem uma estequiometria similar, sugerindo uma interação similar uréia-vírus entre estes sistemas. A desnaturação do TMV utilizando GndHCl foi mais eficiente que a uréia, apresentando estequiometria de 7,50 mols de GndHCl /mol de subunidade de TMV / Abstract: Viruses are very efficient self-assembly structures, but little is understood about the thermodynamics governing this directed assembly. At higher levels of pressure occurs dissociation of TMV or when pressure is combined with urea, denaturation occurs. For a better understanding of such processes, we investigated the apparent thermodynamic parameters of dissociation and denaturation by assuming a steady-state condition. These processes can be measured considering the decrease of light scattering of a viral solution due to the dissociation process, and the red shift of the fluorescence emission spectra, that occurs with the denaturation process. We determined the urea stoicbiometry considering the equilibrium reaction of TMV dissociation and subunit denaturation, which furnished, respectively, 1.53 and 11.1 mois of urea/mol of TMV subunit. The denaturation and dissociation conditions were arrived in a near reversible pathway, allowing the determination of thermodynamic parameters. Gel filtration HPLC, electron microscopy and circular dichroism confirmed the dissociation and denaturation processes. The calculation of apparent urea stoicbiometry of several other viruses based on spectroscopic results from earlier papers showed that dissociation and denaturation processes follow similar stoichiometrics, suggesting a similar virus-urea interaction among these systems. The TMV denaturation was more efficient in the presence of GndHCl, with stoichiometry of 7.50 mois of GndHCl /mol of TMV subunit / Doutorado / Bioquimica / Doutor em Biologia Funcional e Molecular

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-07-17T15:09:58Z No. of bitstreams: 1 2016 - L?gia Marcondes Rodrigues dos Santos.pdf: 1928089 bytes, checksum: 74cdeac055ec3629545f632b08dd75c4 (MD5) / Made available in DSpace on 2017-07-17T15:09:58Z (GMT). No. of bitstreams: 1 2016 - L?gia Marcondes Rodrigues dos Santos.pdf: 1928089 bytes, checksum: 74cdeac055ec3629545f632b08dd75c4 (MD5) Previous issue date: 2016-11-28 / Beer is the most consumed beverage in the world and Brazil is the third biggest producer just behind USA and China. It is an alcoholic beverage made from malted cereals, water, hops and yeast. To produce beer, water and malt are mixed and heated, what it is called the mashing step which aims to promote gelatinization and the starch hydrolyzation by the enzymes of the malt: ? and ? amylases. Since by using high hydrostatic pressure (HHP) it is possible to realize both processes (gelatinization and hydrolysis), mashing can use this technology without the need of heating and to raise productivity of the brewhouse. The aim of this study was to evaluate the use of HHP on starch hydrolysis and the sugar profile formed. For this, worts (water and malt solution ? ratio 4:1) were treated with HHP in 300, 400 or 500 MPa for 5, 10 and 15 minutes, and as control sample, the same solution was treated with heat (1st step: 52?C / 20 min; 2nd step: 72 ?C / 25 min; 3rd step: 78 ?C / 5 min). After, it was made the sugar profile by high performance liquid chromatography (HPLC), soluble solids content by density and microscopic analysis of gelatinized starch. As results, it was possible to verify a decrease on starch concentration (indication of hydrolysis) and a progressive swelling (indication of gelatinization) of the granules, especially at 500 MPa. The iodine test, which assesses starch presence, showed the same results of microscopy analysis and, on both qualitative assessments, the pressure level was of great influence and only on the iodine test, time seems to have had any influence since it was observed a decrease on starch content as time increased at 500 MPa. The sugar profile revealed five main sugars in all worts: glucose, sucrose, fructose, maltose and maltotriose. The maltose, maltotriose and fructose?s contents of the pressurized worts were similar to the heat treatment (500 MPa / 5 ? 15 min). Fructose and glucose exhibited and increase in most of the pressurizations. Sucrose and soluble solids contents of the pressurized samples were lower than the heated one. Among the pressure levels studied, there was a trend of increase on soluble solids, maltose, maltotriose and glucose?s content as the pressure was raised and generally, pressure influenced all sugar?s contents, except from sucrose. Time had a small effect on sugar?s content in some pressure levels. The variations observed to maltose, maltotriose and soluble solids were adjusted to mathematical models of fourth grade that exhibited a strong influence of pressure without dependence of the time effect. Additionally, the principal component analysis explained the variance with formation of three main clusters: heated wort, 500 MPa / 10 or 15 min wort and the other pressurized worts. The results showed that high hydrostatic pressure was able to promote desirable changes on mashing concerning gelatinization and hydrolysis with formation of desirable sugars, especially at 500 MPa. It suggests that it?s possible to produce wort by HHP providing productivity gains, once HHP takes less time (5 min against 80 min of usual heat process) and less energy consumption. / A cerveja ? a bebida alco?lica mais consumida no mundo, e o Brasil ? o terceiro maior produtor. No processo de produ??o convencional, a mostura ? a etapa que objetiva prioritariamente, promover gelatiniza??o e hidr?lise enzim?tica do amido pelas enzimas end?genas ? e ? amilases. A alta press?o hidrost?tica (APH) ? capaz de promover a gelatiniza??o de amido e ativa??o de enzimas e, desta maneira, a mostura demonstra ser uma etapa da fabrica??o de cerveja capaz de utilizar APH, no entanto h? poucos estudos na literatura sobre esta aplica??o. A adequa??o dessa tecnologia poder? propiciar esta etapa do processo sem necessidade de calor e com tempo reduzido, o que reverter? em aumento de produtividade e capacidade da ind?stria cervejeira. Desta maneira, o objetivo deste trabalho foi avaliar o efeito do processo da APH sobre a hidr?lise do amido para forma??o de a??cares durante a mostura. Para isso, mostos (solu??o contendo ?gua e malte ? 4:1) foram tratados por alta press?o hidrost?tica (APH) a 300, 400 ou 500 MPa por 5, 10 ou 15 minutos e, como controle do experimento, a mesma solu??o foi tratada termicamente (TT: 1? fase: 52?C / 20 min; 2? fase: 72 ?C / 25 min; 3? fase: 78 ?C / 5 min) . Posteriormente, procedeu-se an?lise do perfil de a??cares por cromatografia l?quida de alta efici?ncia (HPLC), o teor de s?lidos sol?veis por densitometria e an?lise microsc?pica da gelatiniza??o do amido. Como resultado, observou-se atrav?s da microscopia que houve diminui??o na concentra??o de amido (indicativo de hidr?lise) e um progressivo inchamento (indicativo de gelatiniza??o) dos gr?nulos com o aumento da press?o, principalmente a 500 MPa. O resultado do teste iodo (que avalia presen?a de amido) corroborou com o resultado da microscopia, indicando a diminui??o na concentra??o de amido a 500 MPa. O perfil de a??car revelou cinco sacar?deos principais: sacarose, frutose, glicose, maltose e maltotriose. As concentra??es de maltose, maltotriose e frutose dos mostos pressurizados foram similares ?s observadas no mosto TT quando nos n?veis mais intensos (500 MPa / 5, 10 e 15 min). Frutose e glicose exibiram aumento de concentra??o na maioria dos tratamentos avaliados. O teor de sacarose e de s?lidos sol?veis foram inferiores ao observado no mosto TT em todos os mostos tratados por APH. Entre os n?veis de APH avaliados, observou-se tend?ncia de aumento da concentra??o de s?lidos sol?veis, maltose, maltotriose e glicose com o aumento da intensidade de APH. De maneira geral, o efeito da press?o alterou o teor de todos os a??cares, exceto sacarose. J? o tempo exerceu discreto efeito na concentra??o dos a??cares quando pressurizados em alguns n?veis de press?o. O modelo matem?tico que melhor se ajustou aos dados experimentais dos teores de maltose, maltotriose e s?lidos sol?veis foi o de quarto grau, que exibiu forte influ?ncia do fator press?o sem depend?ncia do fator tempo, corroborando com as outras an?lises deste estudo. Adicionalmente, an?lises de componentes principais exibiu explica??o (CP1 + CP2) a 92 % de vari?ncia com forma??o de tr?s clusters principais: mosto TT, mostos 500 MPa / 10 ou 15 min e demais mostos tratados por APH. Os resultados demostraram que a APH foi capaz de promover as altera??es desej?veis no processo de mostura no que se refere aos efeitos de gelatiniza??o e hidr?lise com forma??o dos a??cares de interesse, principalmente nos n?veis mais intensos (500 MPa) de press?o. Esse resultado sugere que a produ??o de mosto por APH ? vi?vel tecnicamente, podendo vir a proporcionar ganho expressivo de produtividade, uma vez que a APH demanda menos tempo (5 min contra 80 min do tratamento t?rmico) e h? redu??o do consumo de energia.

La mise en œuvre des hautes pressions permet d'inactiver des micro-organismes contaminants (levures, bactéries, spores, virus…) au sein de divers milieux. Cette technologie a tout d'abord été utilisée puis développée à l'échelle industrielle pour décontaminer des produits alimentaires. Du fait de la faible énergie générée par les hautes pressions, seules les liaisons faibles et correspondant à une valeur de ?V négative sont altérées, ce qui, en outre, ouvrait la voie à de nombreuses applications biologiques. L'objectif de cette thèse était d'inactiver l'ensemble des micro-organismes pathogènes au sein d'un produit sanguin : le plasma. L’enjeu principal était d'une part de sauvegarder la fonctionnalité des principales protéines contenues dans le plasma (facteurs de coagulation notamment) et d'autre part d'inactiver les micro-organismes contaminants. Afin d’atteindre cet équilibre, plusieurs paramètres ont été utilisés : - la valeur de la pression (de 100 à 300 MPa), - la durée du traitement, - les températures négatives… Plusieurs résultats originaux seront présentés, notamment en ce qui concerne la différence de sensibilité aux hautes pressions des protéines d'une part et l'inactivation de micro-organismes d'autre part. / High hydrostatic pressure (HHP) has shown its capacity to inactivate contaminating micro-organisms (yeasts, bacteria, spores, viruses…) present in various media. This technology has been first used and developed to an industrial scale for the food industry. Because of the low energy raised using high pressure, only weak interactions characterized by a negative ?V will be altered. This statement opens the way to numerous applications in the biology field. The objective of this PhD work was to inactivate all the pathogens that may contaminate blood products, and blood plasma in particular. The main challenge was to inactivate the contaminating micro-organisms while maintaining the activity of the plasma proteins (such as blood coagulation factors). To achieve this balance, several parameters were used: - the pressure value (from 100 to 300 MPa), - the treatment duration, - negative temperatures… Some original results will be presented, concerning differences in the high pressure sensitivity of proteins and the inactivation of pathogens.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20264号 / 医博第4223号 / 新制||医||1021(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 椛島 健治, 教授 松田 秀一, 教授 羽賀 博典 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Foodborne disease outbreaks associated with the protozoan parasite Cryptosporidium spp. are an emerging public health concern. Shellfish, including Eastern oysters (Crassostrea virginica) in Chesapeake Bay and other Atlantic coastal waters, have been identified as a potential source of Cryptosporidium parvum infection for humans. The inactivation of C. parvum and other pathogens in raw molluscan shellfish would provide increased food safety for normal and at-risk consumers. The objectives of this study were to evaluate the efficacy of three alternative food-processing treatments (e-beam irradiation, microwave energy, and high hydrostatic pressure processing) on the viability of C. parvum oocysts in Eastern oysters. Oysters were artificially infected with the Beltsville strain of C. parvum and subjected to the three treatments in separate experiments. The effects of the treatments were evaluated by inoculation of the processed oyster tissues using the neonatal mouse bioassay. E-beam radiation of in-shell and shucked oysters treated with doses of 1.0, 1.5 or 2.0 kGy produced significant reductions (P < 0.05) in C. parvum mouse infectivity. A dose of 2.0 kGy completely terminated the infectivity of C. parvum and did not adversely affect the visual appearance of the oysters. Microwave treatments of shucked oysters at time (temperature) exposures of 1 sec (43.2°C), 2 sec (54.0°C), and 3 sec (62.5°C) produced a reduction in C. parvum mouse infectivity of 26.7%, 33.3%, and 46.7%, respectively. Microwave treatments at 2 sec (54.0°C) and 3 sec (62.5°C) showed extensive changes in oyster meat texture and color. Thus due to lack of efficacy and unacceptable tissue changes, microwave treatment of oysters is not considered a viable food processing method. High pressure processing of shucked oysters at all pressures tested (305 MPa, 370 MPa, 400 MPa, 480 MPa, 550 MPa) significantly (P < 0.05) reduced infectivity of C. parvum oocysts as measured by the neonatal mouse bioassay. A treatment of 550 MPa at 180 sec produced the maximum decrease of C. parvum infectivity (93.3%). The results indicate that HPP (high pressure processing) can produce significant (P < 0.05) reductions in infectivity of C. parvum oocysts. Measurement of tristimulus color values of pressurized raw oysters at extended processing times from 120 sec to 360 sec at 550 MPa showed an increase (P < 0.05) in whiteness. One (e-beam irradiation) of the three food-processing treatments shows promise for commercial applications to reduce public health risks from cryptosporidiosis in Eastern oysters. / Ph. D.

The Food and Drug Administration has recently expressed concern for the safety of seafood and seafood products. One of the concerns is the presence of Bacillus cereus in fresh blue crab meat. Bacillus cereus is a spore-forming pathogen whose spores survive the customary thermal treatments applied during cooking and pasteurization; therefore it could potentially present a health concern to consumers as the microorganism could increase to pathogenic levels. The objectives of this study were to evaluate the effects of a post-processing method i.e. high hydrostatic pressure treatment on the quality of fresh crab meat and to evaluate the effectiveness of high pressures on the inactivation of B. cereus spores. Fresh blue crab meat was pressurized at 300 and 550 MPa at 25° C for 5 min and stored at 4° C for 31 days to determine the pressurization effects on the microbiological, physical, and sensory quality of the meat. A pressure of 300 MPa caused a 1 log reduction in total aerobic plate count and a 3 day lag period, whereas 550 MPa inactivated 2 logs in total aerobic plate count with no evident lag phase. Physical and sensory qualities of pressurized crab meat were not statistically different from the untreated crab meat (P>0.05). A pressure of 300 MPa extended the shelf-life from 17 to over 24 days with the prevalence of Carnobacterium piscicola at the time of spoilage. Crab meat treated with 550 MPa was not rejected by sensory panels at day 31 and Enterococcus spp. was identified as the predominant microorganism. High hydrostatic pressure (550 MPa at 40° C for 15 min) inactivated less than 1 log (0.66 log) of B. cereus spores inoculated in fresh crab meat. The meat essentially had a protective effect on pressure inactivation of the spores. During storage (31 days), surviving B. cereus was suppressed and outgrown by the other pressure resistant microflora at a storage temperature of 12° C. At 4° C, B. cereus could compete with the other pressure-resistant microflora and was isolated even at the end of the storage period (day 31); however, diarrheal toxin was not detected in any stored samples. / Ph. D.

京都大学 / 新制・論文博士 / 博士(医学) / 乙第13634号 / 論医博第2325号 / 新制||医||1074(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 尾野 亘, 教授 伊達 洋至, 教授 安達 泰治 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM