Submitted by William Justo Figueiro (williamjf) on 2015-07-20T22:41:53Z No. of bitstreams: 1 56c.pdf: 3770514 bytes, checksum: cf71b027cc3d4c64d9a8799069053dd1 (MD5) / Made available in DSpace on 2015-07-20T22:41:53Z (GMT). No. of bitstreams: 1 56c.pdf: 3770514 bytes, checksum: cf71b027cc3d4c64d9a8799069053dd1 (MD5) Previous issue date: 2014-07-31 / Nenhuma / As culturas tradicionais de bactérias lácticas utilizadas para elaboração dos leites fermentados pós-acidificam mesmo em temperatura de refrigeração. Uma seleção cuidadosa poderia reduzir os impactos causados pelas falhas na cadeia do frio e melhorar a qualidade sensorial dos produtos oferecidos ao mercado consumidor. Diante disso, o objetivo deste trabalho foi selecionar culturas microbianas adequadas para o laticínio, sendo que a principal característica buscada foi baixa pós-acidificação e aumento de viscosidade do iogurte e da bebida láctea. Foram avaliadas as três culturas atualmente utilizadas pelo laticínio e cinco novas culturas para os produtos bebida láctea e iogurte. Para ambas as culturas foi verificado o tempo de fermentação, a pós-acidificação quando o produto era mantido em temperatura de fermentação, além do pH e viscosidade durante o shelf life, quando o produto era resfriado à 10 e 15 °C antes do envase. Os resultados mostraram elevada pós-acidificação e baixa viscosidade para duas culturas atualmente utilizadas pela empresa, as quais foram descontinuadas. Três novas culturas apresentaram resultados satisfatórios para o objetivo proposto. Foi possível concluir que a escolha da cultura microbiana interfere fortemente na pós-acidificação e viscosidade para o produto bebida láctea e que a interferência não é tão significativa para o produto iogurte. Através da realização deste trabalho foi verificado que o shelf life da bebida láctea poderia ser aumentado de 45 para 60 dias. / The traditional cultures of lactic acid bacteria used in the preparation of fermented milk post-acidify even at refrigeration temperature. A careful selection could reduce the impacts caused by failures in the cold chain and improve the sensory quality of products offered to the consumer market. Therefore, the aim of this study was to select appropriate microbial cultures for the dairy industry, and the main feature was sought low post acidification and viscosity increase of fermented milks. The three cultures currently used in the dairy industry and five new cultures for yogurt and fermented dairy drink products were evaluated. Fermentation time was checked for both cultures, when the after acidification from the product was maintained at a temperature of fermentation, addition of pH and viscosity during the shelf life, when the product was cooled at 10 and 15 º C before packaging. The results show low viscosity and high post acidification for two currently cultures used by the company, which were discontinued. Three new cultures showed satisfactory results for the proposed objective. It was concluded that the choice of the microbial culture interferes strongly in the post acidification and viscosity for the product fermented dairy drink and that interference is not as significant for the yogurt product. Through this study it was found that the shelf life of fermented milk could be increased from 45 to 60 days.

L’acide hyaluronique (AH) est un polymère osidique aux propriétés mécaniques et biologiques d’intérêt pour le secteur de la santé et des cosmétiques. L’AH est produit à l’échelle industrielle par mise en oeuvre de souches de streptocoques groupe C et D. A l’issue de la production, une part de l’AH est libre dans le milieu de culture complexe, l’autre est associée à la capsule des cellules. La purification de l’AH est délicate en raison, d’une part, de la complexité des milieux de production et, d’autre part, des exigences de la pharmacopée européenne. L’objectif de cette thèse a été de proposer un enchaînement d’opérations et de conditions opératoires permettant une purification de l’AH à partir d’un milieu de production. Au départ, deux méthodes analytiques originales de quantification et d’évaluation de la taille de l’AH en milieux complexes ont été mises au point afin de pouvoir évaluer les critères de performance des procédés de séparation. Ensuite, quatre opérations permettant, premièrement l’extraction, deuxièmement l’élimination des cellules productrices d’AH, troisièmement la purification, et enfin, le séchage d’AH ont été établies. L’étude de l’extraction de la capsule d’AH assistée par SDS ou au TCA a permis d’établir que l’AH lié aux cellules ne représente que 7 % de l’AH produit. Il en a été déduit que cette étape n’est pas nécessaire. L’étude de la séparation des cellules du milieu de culture a montré que la microfiltration tangentielle n’est pas adaptée en raison, d’une part, d’une rétention trop élevée de l’AH (même avec des diamètres de pores supérieurs à 0,22 µm) et d’autre part, d’un colmatage trop rapide des membranes. Cette étape doit être réalisée par filtration frontale sur adjuvant de filtration (Célite). La purification de l’AH a été envisagée par diafiltration (DF) avec des membranes d’ultrafiltration. L’étude des conditions opératoires (PTM, débit d’alimentation, concentration en AH et seuil de coupure), par plan d’expériences, a permis de sélectionner des conditions qui maximisent le flux de perméat ainsi que la rétention d’AH. Ces conditions ont été appliquées en diafiltration au cours de laquelle les performances de séparation (rendement, pureté, productivité) ont été étudiées. Une méthodologie, basée sur les équations classiques de bilans de matière et de flux de perméat en DF, a alors été proposée. Cette démarche avait tout d’abord pour but de prédire lesdites performances en prenant en compte la composition complexe du milieu, de culture bactérienne mais aussi de faciliter l’agencement des étapes du procédé de purification. La diafiltration permet de satisfaire les exigences de pureté de la pharmacopée, mais pas les niveaux de certaines molécules (protéines et acides nucléiques). L’étude de l’adsorption de ces molécules sur charbon actif, en fonction de la température, de la force ionique, du pH et du type de charbon actif, a permis d’établir les conditions les plus favorables de l’élimination de ces molécules. Les équations classiques de modélisation des isothermes correspondant le mieux aux résultats expérimentaux ont été sélectionnées et utilisées avec les équations de performance en DF, afin d’associer au mieux ces deux étapes. Enfin, concernant le séchage de l’AH, deux méthodes, l’une par lyophilisation et l’autre par précipitation en solvant organique, ont été comparées. Ces dernières permettent d’obtenir moins de 20 % d’humidité et de conserver les propriétés des molécules d’AH / Hyaluronic acid (HA) is a biopolymer with mechanical and biological properties interesting the health and cosmetic areas. The industrial HA production is realized by implementation of streptococcus strains (group C or D). At the end of the production phase, a part of HA is free in a complex culture medium while the other remains associated to the cells in a capsule. The two main difficulties of the purification of HA lie, firstly, on the complexity of the production medium and, secondly, on the high requirements of the European Pharmacopoeia for such an application. The aim of this thesis was to provide an appropriate sequence of separation processes and a set of operating conditions for HA purification from a complex production medium which complies with the Pharmacopea. Initially, two original analytical methods for quantifying and evaluating the size of HA in complex environments have been developed in order to evaluate the performance criteria of the separation processes. Then, the HA extraction from the capsule, the elimination of cells, the purification and drying of HA were using classical separation processes and studied. The HA extraction by SDS or TCA showed that the HA bound to cells corresponds to 7 % of HA production. As a consequence, it was concluded that this step was not necessary. The cell separations from the culture medium by tangential microfiltration was unsuitable because of an excessive retention of HA (even at pore diameters higher than 0.22 µm) and a too fast membrane clogging. Good performances were observed with dead-end filtration using Celite. Then, the purification of HA by diafiltration (DF) with ultrafiltration membranes was considered. The operating conditions (PTM, feed rate, HA concentration and cutoff) allowing to maximize the permeate flow rate and the HA retention were selected from design of experiments methodology. The performances of purification by diafiltration (DF) in these conditions (yield, purity and productivity) were monitored. A method for predicting the performances according to the complex medium composition, based on the conventional mass balance equations and the DF permeate flow rate, was then proposed. This was done in order to facilitate the arrangement of the purification steps. In DF, the requirement of the pharmacopoeia in term of overall purity was obtained (> 95 %) but the proteins and nucleic acids levels were still above the threshold. This is why the adsorption of these molecules on activated carbon was studied as a function of temperature, ionic strength, pH and the type of activated carbon. The isotherm equations that better fitted the experimental data were selected and used combined to the equations of DF performances in order to find an appropriate chaining. Finally, the AH lyophilization and precipitation in organic solvent were compared. Both drying methods present less than 20 % moisture and preserve the properties of the molecule

Användandet av deponier är en av de vanligaste metoderna för avfallshantering globalt. Trots insatser som gjordes för att förbjuda hushållsavfall i deponier under millennieskiftet, deponier skapade innan restriktionerna är fortfarande en risk för miljön. Under 2014 öppnade SÖRAB en kontinuerlig biologisk reningsanläggning (KBR-anläggning) på Löt Avfallsanläggning för att hantera lakvatten från en gammal deponi som under en tid fylldes med hushållsavfall. Sedan dess har SÖRAB arbetat med att förbättra KBR-anläggningen. Målet med denna studie är att utforma en driftstrategi för KBR-anläggningen för att förbättra kvävereningen vid låga temperaturer. Ett antal laborativa försök genomfördes, såsom den mikrobiella konsortiets livsduglighet i lakvattnet och tillväxten i både rumstemperatur och vid 4°C, bioaugmentation genom att berika den mikrobiella cellkulturen som redan finns i lakvattnet och hur detta förbättrar kvävereningen i jämförelse med tillsatser av den kommersiella bakterieblandningen ClearBlu Environmental och andra externa kolkällor. Resultaten från dessa laborativa försök påvisade komplett nitrifikation i både rumstemperatur och 4°C i berikat lakvatten från KBR-anläggningens L2A bassäng efter fem dagar. Försöket visade även på syresatt denitrifikation. Dessutom påvisades komplett denitrifikation inom fem dagar, vid rumstemperatur i lakvatten från anläggningens L2B bassäng. Under efterföljande pilotförsök påvisades möjligheten till upplivandet av den biologiska kvävereningen genom berikningen av den mikrobiella cellkulturen i lakvattnet. I ett pilotförsök då lakvatten från L2B bassängen berikades, komplett denitrifikation skedde under en anaerob fas på 16 dagar samt nitrifikation och aerob denitrifikation under ett påföljande 17 dagar lång aerob fas. Ett annat pilotförsök då lakvatten från L2A bassängen berikades påvisade både aerob och anaerob nitrifikation, då ammoniumrening skedde i både den syresatta och syrefria fasen. Tillsatsen av nutrient broth (näringsbuljong) kan påverka KBR-anläggningen, vilket kväver vidare studier. Resultatet från detta projekt tydligt påvisar att kvävereningen i KBR-anläggningen kan förbättras genom att berika den redan närvarande mikrobiella kulturen. / Landfilling has been one of the most popular methods of handling waste globally. Despite the efforts made to stop the disposal of household waste during the turn of the millennia, the landfills formed before these restrictions are still at risk for causing harm to the environment. In 2014, SÖRAB opened a continuous-flow biological treatment (KBR) facility in Löt to treat the leachate produced in one of their older landfills, once filled with household waste. Since then, SÖRAB has been working on improving the treatment facility. The aim of this the study is to find a suitable process to enhance the nitrogen removal at low temperature. Several laboratory scale experiments were performed, such as viability of microbial consortia in the leachate and growth at room temperature and at 4°C, testing bioaugmentation by enriching the microbial cell culture in the leachate and their efficiency in removing nitrogen, compared to the commercial cell culture ClearBlu Environmental and carbon source addition. The results displayed complete nitrification at both room temperature and 4°C in bioaugmented, enriched leachate originating from the L2A basin of the KBR facility, after five days. These trials also suggested the occurrence of aerated denitrification. Complete denitrification within five days was seen at room temperature in bioaugmented, enriched leachate from the L2B basin of the same facility. The ensuing pilot scale trials proved the possibility to revive the biological nitrogen removal by microbial cell culture enrichment. In one pilot in which leachate from the L2B basin was enriched, complete denitrification in the anaerobic phase consisting of 16 days occurred, along with some nitrification and aerated denitrification in the 17 day long aerated phase that followed. Another pilot scale trail in which leachate from the L2A basin was enriched, both aerobic and anaerobic nitrification occurred, as ammonium removal occurred in both the aerated and unaerated phases. The addition of nutrient broth might influence the KBR system which needs further study. The results from this project clearly demonstrate that nitrogen removal in the KBR facility could be enhanced using a culture naturally present in the facility.