Superheated steam (SS) drying of distillers’ spent grain (DSG) is a more energy efficient alternative to conventional hot air drying. SS drying at sub-atmospheric pressure (also referred to as low pressure) can prevent burning and lowering the quality of the food product. The objective of this study was to design, fabricate, and test a SS drying system that could operate at sub-atmospheric pressure for drying DSG. After the custom designed system was constructed, major problems associated with the system were identified. A number of tests were carried out and modifications were made to the system to resolve technical problems. Distillers’ spent grain was then successfully dried using the system under various levels of temperature from 95 to 115°C and pressure of either -25 or -20 kPa, with a SS velocity from 0.100 to 0.289 m/s.

Biomass such as spent grain is difficult to dry when it is in the slurry form. Proposed industrial solutions are to compact wet biomass first and then dry it. Compaction develops desired granular form and increases surface area for drying but also brings new technical challenges. Superheated steam (SS) drying is advantageous over hot-air drying as it is more energy efficient. A problem associated SS drying is the initial condensation leading to disintegration of biomass compacts. The current research investigates the disintegration characteristics of distiller’s spent grain (DSG) compacts while being dried in SS. The study focuses on the DSG flowability, densification characteristics and disintegration behavior of DSG compacts as affected by SS drying conditions, soluble content and particle size distribution (PSD). DSG fractions with particle sizes from 300 to 850 µm were dried in SS at 150°C and hot-air at 45 and 150°C. Under these drying conditions bulk density and angle of repose (AOR) varied from 0.379 to 0.435 g/cm3 and 46.0 to 50.4°, respectively. The stress-relaxation data obtained during the compaction of DSG at different levels of compressive pressure (60.3-135.7 MPa), initial moisture content (15, 20 and 25% wet basis- wb) and soluble content (15 and 30%) were normalized and analyzed to determine the asymptotic modulus (EA) of the compacts. The highest EA of 174 MPa was obtained for DSG compacts produced with a compressive force of 135.7 MPa, initial moisture of 25% wb and soluble content of 0%. The percentage increase in volume of DSG compacts during drying in SS at 110 to 150°C temperature range was between 78 to 130%. A comparison between the physical properties of SS dried and hot-air dried compacts revealed the role of SS in accelerating the release of mechanical energy stored in the compacts. An increase of dimensions and a considerable increase in the hardness and EA of the compacts was obtained by adding up to 70% (w/w) solubles or by decreasing the PSD of wet distiller’s spent grain from d(0.9)=1283.6 to 812.8 µm. This study establishes that compaction of wet biomass followed by SS drying can lead to its effective utilization.

Superheated steam (SS) drying of distillers’ spent grain (DSG) is a more energy efficient alternative to conventional hot air drying. SS drying at sub-atmospheric pressure (also referred to as low pressure) can prevent burning and lowering the quality of the food product. The objective of this study was to design, fabricate, and test a SS drying system that could operate at sub-atmospheric pressure for drying DSG. After the custom designed system was constructed, major problems associated with the system were identified. A number of tests were carried out and modifications were made to the system to resolve technical problems. Distillers’ spent grain was then successfully dried using the system under various levels of temperature from 95 to 115°C and pressure of either -25 or -20 kPa, with a SS velocity from 0.100 to 0.289 m/s.

O presente estudo teve como objetivo avaliar a produção de etanol pela levedura Pichia stipitis, a partir do hidrolisado hemicelulósico do bagaço de malte (HHBM). Primeiramente estudou-se o efeito da suplementação nutricional do hidrolisado adicionando-se extrato de farelo de arroz (0 a 20% v/v), uréia (0 a 3 g/L) e extrato de levedura (0 a 3 g/L). Os resultados mostraram que o hidrolisado suplementado apenas com extrato de levedura proporcionou os melhores resultados de conversão (YP/S = 0,44 g/g) e produtividade em etanol (QP = 0,33 g/L.h). Em seguida foi avaliado o nível ótimo deste nutriente sobre a bioconversão, sendo confirmada a suplementação do HHBM com 3,0 g/L de extrato de levedura. Após o estabelecimento das condições nutricionais, a levedura foi cultivada em HHBM após o crescimento de 24 horas em meio semissintético, visando aclimatar as células aos inibidores presentes no hidrolisado. Este estudo resultou no aumento das velocidades do processo, sendo 16% sobre a produtividade volumétrica em etanol (QP) e 10% sobre a velocidade de consumo de substrato (QS), quando comparado ao cultivo das células somente em meio semissintético. Na etapa seguinte utilizou-se um planejamento fatorial 22 com face centrada para otimização das condições de pH e concentração inicial de células (X0) na fermentação em HHBM não destoxificado e após a destoxificação com carvão ativado. Foram obtidos modelos para descrever os valores de YP/S e QP na região estudada, sendo que para o HHBM não destoxificado os valores destes parâmetros foram 0,40 g/g e 0,65 g/L.h, respectivamente, em pH 6,4 e X0 = 5,0 g/L (modelo 1). Em HHBM destoxificado, as condições ótimas foram obtidas em pH 6,0 e 1,36 g/L de células (modelo 2), obtendo-se YP/S de 0,40 g/g e QP de 0,46 g/L.h. Nas condições otimizadas foram então realizados ensaios que confirmaram a validade dos modelos 1 e 2, obtendo-se a máxima concentração de etanol (23,4 g/L), YP/S de 0,41 g/g e QP de 0,65 g/L.h em HHBM não destoxificado. Realizouse ensaios para avaliação do efeito do ácido acético sobre a fermentação em meio semissintético por P. stipitis, empregando-se as condições de pH e X0 otimizadas em HHBM não destoxificado (modelo 1) e destoxificado (modelo 2). Este estudo mostrou que nas condições do modelo 1, o ácido acético favoreceu a bioconversão sendo os melhores resultados obtidos na presença deste ácido (YP/S = 0,47 g/g e QP = 1,08 g/L.h). Por outro lado, nas condições do modelo 2, os valores de YP/S foram similares, enquanto que com a adição de ácido acético ao meio de fermentação, o valor de QP foi reduzido em 53%. Na fermentação em biorreator, o emprego das condições otimizadas em frascos (pH 6,4 e 5,0 g/L de células) resultaram em valores de QP 48% inferiores ao obtido em frascos (0,65 para 0,44 g/L.h), entretanto YP/S foi apenas 10% inferior (0,41 para 0,37 g/g). No presente estudo, conclui-se que a suplementação nutricional do HHBM e a otimização das condições de pH e X0 resultaram em valores promissores para os principais parâmetros da fermentação por P. stipitis, ressaltando o potencial deste hidrolisado em processos biotecnológicos para produção de etanol. / This study aimed to evaluate the ethanol production by Pichia stipitis in brewer\'s spent grain hemicellulosic hydrolysate (BSGHH). Initially, the effect of nutritional supplementation was evaluated by adding rice bran extract, urea and yeast extract. The results showed that supplementation only with yeast extract promoted the highest conversion values (YP/S = 0.44 g/L) and ethanol productivity (QP = 0.33 g/L.h). Additional assays showed that the optimal concentration of this nutrient was 3.0 g/L. To acclimate the cells to inhibitors present in BSGHH the yeast was cultivated in hydrolysate after growth for 24 hours in semisynthetic medium. This study resulted in increased of the process rates, 16% of the ethanol productivity (QP) and 10% on the substrate consumption (QS) when compared to growing cells only in the semisynthetic medium. In the second step a 22 full factorial centeredface design was employed to optimized the conditions of pH and initial cells concentration (X0) in fermentation of hydrolysate undetoxified and after detoxification with activated charcoal. Mathematical models that relate the YP/S and QP were obtained. For non-detoxified BSGHH (model 1) the optimal conditions of pH (6.4) and X0 (5.0 g/L) showed values parameters of 0.41 g/g and 0.65 g/L.h, respectively. In detoxified BSGHH (model 2) the optimum conditions of pH (6.0) and X0 (1.36 g/L), resulted in YP/S and QP values of 0.40 g/g and 0.46 g/L.h, respectively. Under these conditions, the the validity of the models were confirmed. The effect of acetic acid on fermentation by P. stipitis in semisynthetic medium, employing optimized conditions of pH and X0 in model 1 and model 2 was evaluated. The results showed that under the conditions of model 1 and in a concentration of 2,9 g/L, the acetic acid favored the bioconversion by P. stipitis, increasing the YP/S (15 %) and QP (66 %). On the other hand, in the conditions of the model 2 the YP/S values were similar, whereas the QP values were reduced by 53% when the acetic acid was added. By using these optimized conditions in bioreactor fermentation it was obtained the ethanol productivity was approximately 48% lower (0.65 to 0.44 g/L.h), however the ethanol production was similar as compared to fermentation flasks. It is possible conclude that the HHBSG requires nutritional supplementation and that the optimized conditions of pH and initial cells concentration can be used as a strategy in order to raising the fermentation parameters.

Unwanted uranium released in the aquatic environment from uranium mining and nuclear fuel industry has become a growing threat to human health and environment safety due to its radiological and chemical toxicity. Biosorbents from agro-industrial waste are the most preferred materials for the removal of uranium from the wastewater due to their good cost-to-performance ratio. Brewer’s spent grain (BSG), a widely produced by-product from the beer brewery industry, is an inexpensive and readily available feedstock for the production of uranium biosorbents. In the current work, the use of BSG as a promising starting feedstock for low-cost and efficient adsorbents with high adsorption capacity, fast kinetics, selectivity, and reusability, is investigated. Functionalization methods such as thermal treatment, chemical modification (oxidation), and polymer grafting were explored, and the selectivity was tuned using surface ion-imprinting technology. The adsorption performance of adsorbents prepared from BSG was tested under various conditions for practical application, and structure affinity principles were derived from the characterization, data modeling and experimental results (Fig. 1). In the first part of this work, BSG is successfully converted into altered BSG (ABSG), an effective biosorbent, by mild hydrothermal treatment approach (150 ℃, 16 h). Compared with the conventional hydrothermal carbonation method (up to 250 ℃), the current method is carried out at a significantly lower temperature without any additional activation process, which minimizes the energy consumption and environmental impact during the treatment. Maillard reaction plays an important role in increasing the adsorption capacity by forming various Maillard reaction products (methylglyoxal-derived hydroimidazolone-1 with the highest content) and melanoidins with a large number of functional groups. In addition, other pathways such as dehydration, decarboxylation, aromatization and oxidation also contribute to the increased adsorption capacity. Therefore, the content of carboxyl groups in ABSG increases up to 1.46 mmol/g with maximum adsorption capacities for La(Ⅲ), Eu(Ⅲ), Yb(Ⅲ) (pH = 5.7), and U(Ⅵ) (pH = 4.7) of 38, 68, 46 and 221 mg/g, respectively (estimated by the Langmuir model). Moreover, FT-IR spectra show that both O- and N-containing functional groups are involved in the adsorption of studied ions. The second part of this work demonstrates for the first time the successful oxidization of BSG using 85 wt% H3PO4 and NaNO2, increasing the carboxyl groups content from 0.15 mmol/g for BSG to 1.3 mmol/g for oxidized BSG (OBSG). OBSG exhibits fast adsorption kinetics in 1 h and an adsorption capacity for U(Ⅵ) of 297.3 mg/g (c0(U) = 900 mg/L, pH = 4.7), which is superior to other biosorbents reported in the literature. Possible adsorption mechanisms are based on ion-exchange between UO22+ and H+ released from carboxyl groups, and the complexation of UO22+ with the two oxygen atoms of carboxyl groups. For practical application, adsorption/desorption studies show that OBSG retains 60% of original adsorption capacity (167 mg/g) with a desorption ratio of 89% after 5 adsorption/desorption cycles. Evaluation of OBSG performance in simulated seawater (10.8 mg/g, c0(U) = 10 mg/L, 193 mg/L NaHCO3 and 25.6 g/L NaCl, pH0 = 7.7) indicates a potential usage at low concentration, high salinity, and in the presence of carbonate. In the third part of this work, brewer’s spent grain supported superabsorbent polymers (BSG-SAP) with various cross-linking density are prepared for the first time via one-pot swelling and graft polymerization of acrylic acid (AA) and acrylamide as low-cost and environmentally friendly adsorbents. A 7 wt% NaOH solution was used as a swelling agent for BSG and as a neutralization agent for AA without generating alkaline effluents. The use of BSG and graft polymerization can significantly increase the available hydroxyl, carboxyl and amide groups, resulting in a highly cross-linked and highly hydrophilic three-dimensional polymer network of BSG-SAP. The BSG-SAP (BSG-SAP-H) prepared with high cross-linking density exhibits better properties with exceptional adsorption capacity for U(VI) of 1465 mg/g (estimated by the Toth model) at pH0 = 4.6 within 45 min. It also shows good selectivity for U(VI) in the presence of several metal ions (V(V), K(I), Na(I), Mg(II), Zn(II), Co(II), Ni(II), and Cu(II)) with selectivity coefficients (SU) higher than 72%. In simulated seawater, BSG-SAP-H showed higher adsorption capacity (17.6 mg/g for c0(U) = 8 mg/L, pH0 = 8) compared to the currently reported adsorbents based on natural polymers. In the experiments with the fixed bed column (c0(U) = 30 mg/L), the uranyl ions could be concentrated up to 15 folders in U(VI)-spiked water and up to 13 folds in simulated seawater. Moreover, after four cycles, BSG-SAP-H was able to maintain 80% of adsorption capacity in U(VI)-spiked water (254.4 mg/g) and 90% in simulated seawater (37.4 mg/g). FT-IR and 13C solid-state NMR spectra show the function of amide groups for U(VI) adsorption, the bidentate binding structure between UO22+ and the carboxyl groups, and the cation exchange between Na+ in BSG-SAP and UO22+. The fourth part of this work describes a new strategy for the preparation of surface ion imprinted brewer’s spent grain (IIP-BSG) using binary functional monomers (2-hydroxyethyl methacrylate and diethyl vinylphosphonate) for selective removal of U(VI). A high monomer/template molar ratio of 500:1 is used to ensure high site accessibility and easy template removal. IIP-BSG exhibits a maximum U(VI) adsorption capacity of 165.7 mg/g (pH0 = 4.6, estimated by the Sips model), a high selectivity (SU > 80%) for U(VI) in the presence of an excess amount of Eu(III) (Eu/U molar ratio = 20), and good tolerance to salinity (47.4 mg/g for U(VI) at ionic strength = 1 mol/L and c0(U) = 0.5 mM = 120 mg/L). After 5 adsorption and desorption cycles, IIP-BSG retains 90% of its adsorption capacity (36.9 mg/g) and high selectivity (SU > 92%) in binary U(VI)/Eu(III) solution (c0 = 0.5 mM = 120 mg/L). In addition, FT-IR spectra show the electrostatic interaction and a coordination of uranyl ions by carboxyl and phosphoryl groups, the site energy distribution theory shows the predominant contribution of high-energy (specific) sites during selective adsorption, and the kinetic model shows that the internal mass transfer is the rate-determining step of U(VI) adsorption. In the last part of this work, the additional tests were performed for BSG and its derived adsorbents to evaluate their potential for practical application. BSG and most of its derived adsorbents retain 90% of their adsorption capacity after aging in water for 6 days, except for ABSG (60% decrease in adsorption capacity). IIP-BSG shows efficient separation of U(VI)/Ln(Ⅲ) (e.g. La(III), and Nd(III), Sm(III)) in weakly acidic nuclear wastewater (pH0 = 3.5) and U(VI) concentration in carbonate-rich-mine water (e.g. Schlema mine water, pH0 = 7.1) and tailings water (e.g. Helmsdorf tailings water, pH0 = 9.8), demonstrating a high potential for practical use. Selectivity of IIP-BSG is also given for acidic mine water (e.g. Königstein mine water, pH0 = 2.6). In addition, the unmodified BSG and BSG-SAP-H could effectively remove uranyl ions from acidic mine water with high selectivity. In particular, the cost efficiency and the availability of unmodified BSG make it of great interests for the remediation of uranium containing acidic mine water (Table 1).

Brewer's spent grain (BSG) is the major byproduct generated by the brewing industry, which contains 14–30% protein and 50–70% of fiber. Currently, BSG is predominantly used as low-value cattle feed or buried in landfills, which is a considerable loss of valuable resources, leading to economic loss and environmental problems. Although research has been done on BSG valorization, the studies are limited to producing a single product (e.g., polyphenols, ethanol, or active carbon) and without further utilization of the produced products. Besides, the economic information available about the production of value-added products from BSG is insufficient. The overall goal of this research is to develop an integrated process to convert BSG into value-added protein-rich feedstuff and cellulose absorbent. The objectives of the research detailed here were to 1) develop a process to simultaneously produce protein-rich (PP) and fiber-rich products (FP) from BSG, 2) assess the replacement of fishmeal with PP in shrimp feed, 3) evaluate the economics of the overall process of PP production at a commercial scale, and 4) explore the potential use of cellulose adsorbent obtained from the FP in removing heavy metals from contaminated water. To attain these objectives, BSG was first subjected to a wet fractionation process to produce PP and FP using different chemical/biological treatments, where the effects of sodium hydroxide, sodium bisulfite, and a protease (Alcalase) at different concentrations were investigated. Under the optimized conditions, the produced PP contained 46% protein and less than 1% fiber. The effectiveness of using PP to replace fishmeal at increasing levels (10–70%) was then evaluated by performing shrimp feeding trials. The results showed that up to 50% of fishmeal in shrimp feed can be replaced by PP without affecting shrimp growth and feed utilization. Moving forward, a techno-economic analysis was conducted to evaluate the economic feasibility of the production of PP. The experimental conditions and results were input into the process simulation model for determining the mass and energy flows. For a processing plant with a capacity of 590 t wet BSG per day, the minimum selling price of PP to achieve a 5% return was determined to be $1044/t, lower than the price of fishmeal, indicating that the use of PP to replace fishmeal in shrimp feed could potentially reduce shrimp farming cost. The utilization of FP will further improve the economic feasibility of the fractionation process. FP was sequentially treated by dilute acid, alkali, and bleach to produce purified cellulose fibers, which were then modified by 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO) oxidation to produce a cellulose adsorbent. The feasibility of the adsorbent in removing heavy metals (especially lead and manganese) from contaminated water was then investigated. Based on the results, the produced cellulose adsorbent showed high adsorption capacities for lead (272.5 mg/g) and manganese (52.9 mg/g). Overall, this study demonstrated that BSG can be upcycled into multiple value-added products via an integrated process. The outcomes of this study not only provide a low-cost and sustainable protein source to the aquaculture industry, and provide a novel adsorbent for the water treatment industry, but also offer alternative ways for the brewing industry to manage BSG. / Doctor of Philosophy / Brewer's spent grain (BSG) is the major byproduct generated by the brewing industry. Currently, BSG is predominantly used as low-value cattle feed or buried in landfills due to its high fiber and low protein contents, which is a considerable loss of valuable resources. Besides, raw BSG contains other nutrients and high water content, the inappropriate management of BSG may introduce environmental concerns. Though technologies have been investigated to valorize BSG by extracting protein from it, the process scaled-up is limited by the high drying costs of wet BSG, heavy chemical consumptions, and a large amount of fiber residue. The overall goal of this research is to develop an integrated process to convert BSG into value-added protein-rich feedstuff and cellulose absorbent. In this study, we developed and optimized a process to produce protein and fiber products from wet BSG. The protein content of the produced protein product was doubled and the fiber content was reduced significantly compared with the raw BSG, which lighted the use of the protein product as an alternative to fishmeal. Fishmeal is an expensive aquafeed ingredient, the aquaculture industry is looking for alternatives to replace it. Herein, we investigated the effectiveness of the protein product as an alternative to fishmeal by conducting shrimp trials. A further economic analysis was conducted to evaluate the economic feasibility of the proposed process for protein and fiber production from BSG. In addition, the fiber product was used for producing a cellulose adsorbent to remove heavy metals from contaminated water. Overall, this study demonstrated that BSG can be upcycled into multiple value-added products via an integrated process. The outcomes of this study not only provide a low-cost and sustainable protein source to the aquaculture industry, and provide a novel adsorbent for the water treatment industry, but also provide alternative ways for the brewing industry to manage BSG.

Aquaculture, the cultivation of aquatic organisms in a controlled environment, offers both economic and nutritional benefits to human society. As there is an increased demand to feed a growing human population, many wild-caught fisheries have struggled due to the overexploitation of resources. Currently, production relies heavily on wild-caught fish to produce fishmeal to feed farm-raised fish. The demand for alternative materials in fish feeds has grown rapidly as fishmeal resources have become limited. Antibiotic resistance emergence in aquaculture systems is another area of concern. Reducing antibiotic use via alternate prophylactic measures to increase host health is an essential area of research; modulation of the host intestinal bacterial community via prebiotics is one possibility. Prebiotics refer to non-digestible food ingredients that are thought to stimulate the growth of beneficial bacteria, consequently benefiting host health by indirectly reducing the possibility of bacterial pathogen proliferation. This occurs through various measures such as competition for space and resources. The intestinal bacterial community has a significant impact on a variety of host factors that include host development, physiology, immunity, and nutrient acquisition. In turn, there are multiple factors impacting the bacterial community, including the presence of pathogens and/or antibiotics, environmental conditions, host genetics, and the diet consumed. To promote environmental sustainability and improve production and animal health in aquaculture, a collaboration was created with Anheuser-Busch of the brewing industry and Maltento, a functional ingredient company. With breweries around the globe, Anheuser-Busch produces consistent, food grade byproducts that are safe for human consumption. Two of the most prevalent brewery byproducts are brewer's spent yeast (BSY) and brewer's spent grain (BSG). BSY contains a variety of beneficial nutrients such as proteins, essential amino acids, and carbohydrates. BSG is high in fiber but low in protein; however, black soldier fly larvae can be cultured on BSG to convert the low-value product into insect biomass to be used in fish feed, as insects themselves are full of beneficial lipids and proteins. The objective of the work presented in this thesis was to evaluate the efficacy of using low-value brewery waste products, converted into high-value feed additives, for aquaculture practices. Specifically, the effects of dietary feed additives on the production, health, and intestinal bacterial community of aquaculture-raised rainbow trout were examined. Inadvertently, benefits of the feed additives on fish subjected to chronic and acute thermal stress were also assessed. Overall, the results of the study found that the feed additives did not significantly change the production efficiency of the rainbow trout, though some increase in growth was observed. When subjected to chronic thermal stress conditions, fish fed the experimental diets outperformed those fed the control diet regarding growth parameters. The intestinal bacterial community of the fish was significantly altered from the beginning of the trial compared to the end of the trial, though differences were not attributed to the feed additives. Instead, the resulting intestinal dysbiosis is believed to have stemmed from the physiological response of the fish to thermal stress conditions. When the fish underwent an acute thermal stress event, causing mortality, fish fed three of the five experimental diets were found to have higher survival rates compared to the control. Ultimately, results of this project suggest that the BSY and BSG-fed insect feed additives may have increased the health and robustness of the fish during a period of thermal stress. However, further research under controlled conditions is needed to evaluate if the observed host health benefits can directly be attributed to the feed additives. / Master of Science / Aquaculture refers to the method of rearing aquatic organism such as fish and shellfish under controlled conditions. Within the food industry, aquaculture is one of the fastest growing sectors, and provides important economic and nutritional benefits to humans. Additionally, aquaculture is an important alternative to fisheries that rely on catching fish from the natural environment. Wild-caught fisheries have struggled due to the overfishing, and unfortunately, many aquaculture practices still rely on wild-caught fisheries to produce fishmeal used in feed for carnivorous fish. Research into alternate protein sources to use in fish feed has been on the rise. Additionally, as the emergence of multi-drug resistant bacteria continues to increase, reducing antibiotic use has become a priority across all fields whether it be healthcare or the food industry. Within aquaculture, using alternative prophylactic measures such as prebiotics to increase animal health and disease resistance could lead to the overall reduction of antibiotic use. Prebiotics are non-digestible food ingredients believed to help the beneficial bacteria within the intestinal track to grow. In turn, the increased numbers of beneficial bacteria reduce the possibility of pathogenic bacteria invading and establishing a presence in the intestinal track. The intestinal microbiome refers to the various organisms, such as bacteria, viruses, and fungi, that live commensally within the host digestive tract. The bacterial community within the intestinal microbiome has many important roles, including effects on host development, physiology, immunity, and nutrient acquisition. Many factors also impact the bacterial community, including the presence of pathogens and/or antibiotics, environmental conditions, host genetics, and the diet consumed. To promote environmental sustainability and improve production and animal health in aquaculture, a collaboration was created with Anheuser-Busch of the brewing industry and Maltento, a functional ingredient company. With breweries around the globe, Anheuser-Busch produces consistent, food grade byproducts that are safe for human consumption. Two of the most prevalent brewery byproducts are brewer's spent yeast (BSY) and brewer's spent grain (BSG). These low-value waste products can consequently be converted into high-value feed additives for use in aquaculture. The objective of the work presented in this thesis was to evaluate the effects of BSY and BSG-fed insect dietary feed additives on the production, health, and intestinal bacterial community of aquaculture-raised rainbow trout. Unintentionally, benefits of the feed additives on fish subjected to chronic and acute high-temperature thermal stress were also explored. Overall, the results of the study found that while the feed additives did not significantly increase the growth of the rainbow trout, benefits were still observed. When subjected to chronically high-water temperatures, fish fed the experimental diets outperformed those fed the control diet regarding growth parameters. The intestinal bacterial community of the fish was significantly altered from the beginning of the trial compared to the end of the trial, though differences are not believed to be caused by the feed additives. Instead, the resulting shift in the bacterial community is believed to have stemmed from the stress-response of the fish triggered by high water temperature. When the fish underwent an acute thermal stress event, which caused mortality, fish fed three of the five experimental diets were found to have higher survival rates compared to the control. Ultimately, results of this project suggest that the feed additives may have increased the health and robustness of the fish while undergoing thermal stress. However, further research under controlled conditions is needed to evaluate if the observed host health benefits can be attributed directly to the feed additives.

Atualmente observa-se um aumento significativo no consumo de cervejas com baixo teor alcoólico e cervejas sem álcool. Existe um crescente interesse por este tipo de produto tanto por parte das indústrias cervejeiras como por parte dos consumidores. Este aumento é principalmente devido a questões de saúde e a razões de segurança no trabalho e nas estradas. Além disso, há países onde o consumo de álcool é proibido por lei. Tendo em vista que o consumidor está em busca de novos produtos com características mais próximas possíveis de uma cerveja convencional, o presente trabalho visa analisar uma nova abordagem para a produção de cerveja lager com baixo teor alcoólico, reutilizando o bagaço de malte, um subproduto da indústria cervejeira, como substituinte de parte do malte. Foram obtidas cervejas com as seguintes proporções de bagaço/malte: 0:100, 25/75, 50/50 e 75/25. Nas cervejas com 50 e 75% de bagaço de malte, observou-se uma redução no tempo de fermentação quando comparadas com a cerveja puro malte. A atenuação real de fermentação das cervejas produzidas também foi menor devido a menor fermentabilidade dos mostos. O teor alcoólico das cervejas obtidas em escala de bancada apresentou valores entre 0,665 e 1,615 % (v/v), o que as caracterizam como cervejas de baixo teor alcoólico A cerveja com 25% de bagaço recebeu as maiores notas na análise sensorial, tendo boa aceitação entre os provadores, e foi escolhida para a produção em escala piloto. O perfil de fermentação da cerveja produzida em escala piloto foi similar ao perfil de fermentação da cerveja produzida em escala de bancada. Também foi realizada uma análise sensorial, comparando a cerveja produzida em escala piloto com duas cervejas comercias. A utilização do bagaço de malte como substituinte do malte na fabricação de cervejas com baixo teor alcoólico é uma estratégia viável, produzindo uma bebida com aceitação sensorial semelhante ao de uma cerveja de baixo teor alcoólico comercial. / Nowadays there has been a significant increase in the consumption of beer with low alcohol content and non-alcoholic beers. There is a growing interest on this type of product by both brewers industries and consumers. This increase is mainly due to health issues and safety reasons at work and on the roads. In addition, there are countries where the consumption of alcohol is prohibited by law. Given that the consumer is looking for new products with characteristics as similar as possible of conventional beer, this study aims to analyze a new approach for the production of lager beer with low alcohol content, by reusing brewer\'s spent grain, a by-product of brewing industry, as a substitute of part of the malt. Beers were obtained with the following proportions of bagasse/malt: 0/100, 25/75, 50/50 and 75/25. In beers with 50 and 75% of bagasse, it was observed a reduction in the fermentation time compared with all-malt beer. The Real Attenuation Fermentation of Beer produced was also lower due to lower fermentability of wort. The beers\' alcohol content obtained in laboratory scale showed values between 0.665 and 1.615% (v/v), which characterize them as low-alcohol beers. Beer with 25% of bagasse received the highest scores in sensory analysis, having a good acceptance among the tasters, therefore it was chosen for the production on a pilot scale. The beer\'s fermentation profile produced on pilot scale was similar to the beer\'s fermentation profile produced in laboratory scale. Sensory analysis was also performed, comparing the beer produced in pilot scale with two commercial beers. The use of brewer\'s spent grain as a substitute in the manufacture of beers with low alcohol content is a viable strategy, producing a beverage with sensory acceptance similar to a commercial low-alcohol beer.

Atualmente observa-se um aumento significativo no consumo de cervejas com baixo teor alcoólico e cervejas sem álcool. Existe um crescente interesse por este tipo de produto tanto por parte das indústrias cervejeiras como por parte dos consumidores. Este aumento é principalmente devido a questões de saúde e a razões de segurança no trabalho e nas estradas. Além disso, há países onde o consumo de álcool é proibido por lei. Tendo em vista que o consumidor está em busca de novos produtos com características mais próximas possíveis de uma cerveja convencional, o presente trabalho visa analisar uma nova abordagem para a produção de cerveja lager com baixo teor alcoólico, reutilizando o bagaço de malte, um subproduto da indústria cervejeira, como substituinte de parte do malte. Foram obtidas cervejas com as seguintes proporções de bagaço/malte: 0:100, 25/75, 50/50 e 75/25. Nas cervejas com 50 e 75% de bagaço de malte, observou-se uma redução no tempo de fermentação quando comparadas com a cerveja puro malte. A atenuação real de fermentação das cervejas produzidas também foi menor devido a menor fermentabilidade dos mostos. O teor alcoólico das cervejas obtidas em escala de bancada apresentou valores entre 0,665 e 1,615 % (v/v), o que as caracterizam como cervejas de baixo teor alcoólico A cerveja com 25% de bagaço recebeu as maiores notas na análise sensorial, tendo boa aceitação entre os provadores, e foi escolhida para a produção em escala piloto. O perfil de fermentação da cerveja produzida em escala piloto foi similar ao perfil de fermentação da cerveja produzida em escala de bancada. Também foi realizada uma análise sensorial, comparando a cerveja produzida em escala piloto com duas cervejas comercias. A utilização do bagaço de malte como substituinte do malte na fabricação de cervejas com baixo teor alcoólico é uma estratégia viável, produzindo uma bebida com aceitação sensorial semelhante ao de uma cerveja de baixo teor alcoólico comercial. / Nowadays there has been a significant increase in the consumption of beer with low alcohol content and non-alcoholic beers. There is a growing interest on this type of product by both brewers industries and consumers. This increase is mainly due to health issues and safety reasons at work and on the roads. In addition, there are countries where the consumption of alcohol is prohibited by law. Given that the consumer is looking for new products with characteristics as similar as possible of conventional beer, this study aims to analyze a new approach for the production of lager beer with low alcohol content, by reusing brewer\'s spent grain, a by-product of brewing industry, as a substitute of part of the malt. Beers were obtained with the following proportions of bagasse/malt: 0/100, 25/75, 50/50 and 75/25. In beers with 50 and 75% of bagasse, it was observed a reduction in the fermentation time compared with all-malt beer. The Real Attenuation Fermentation of Beer produced was also lower due to lower fermentability of wort. The beers\' alcohol content obtained in laboratory scale showed values between 0.665 and 1.615% (v/v), which characterize them as low-alcohol beers. Beer with 25% of bagasse received the highest scores in sensory analysis, having a good acceptance among the tasters, therefore it was chosen for the production on a pilot scale. The beer\'s fermentation profile produced on pilot scale was similar to the beer\'s fermentation profile produced in laboratory scale. Sensory analysis was also performed, comparing the beer produced in pilot scale with two commercial beers. The use of brewer\'s spent grain as a substitute in the manufacture of beers with low alcohol content is a viable strategy, producing a beverage with sensory acceptance similar to a commercial low-alcohol beer.

<p>Pivski trop čini približno 85% od ukupnih sporednih proizvoda proizvodnje piva, i dostupan je po veoma niskim cenama tokom čitave godine. Pivski trop ima veliku perspektivu za primenu u biotehnologiji i proizvodnji visoko vrednih proizvoda. Jedna od veoma ekolo&scaron;ki i ekonomski isplativih alternativa je upotreba pivskog tropa u proizvodnji mlečne kiseline, jer se poslednjih par decenija uočava intenzivan rast potražnje za mlečnom kiselinom. Mlečna kiselina je najvažnija hidroksikarbonska kiselina &scaron;iroko rasprostranjena u prirodi, sa velikom primenom u prehrambenoj, farmaceutskoj, tekstilnoj i hemijskoj industriji i industriji prerade kože.<br />Cilj istraživanja ove doktorske disertacije je ispitivanje primene pivskog tropa u proizvodnji mlečne kiseline. Prvo je izvr&scaron;ena optimizacija enzimske hidrolize pivskog tropa u cilju dobijanja &scaron;to je moguće veće koncentacije redukujućih &scaron;ećera neophodne za mlečno-kiselu fermentaciju. Hidrolizat pivskog tropa je dobijen enzimskom hidrolizom dodatkom komercijalnih enzima za razgradnju skroba i celuloze. Parametri čiji je uticaj na efikasnost enzimske hidrolize ispitanu su: pH vrednost, temperatura hidrolize i količina dodatih enzima. Nakon &scaron;to su određeni najbolji uslovi razgranje pivskog tropa, dobijeni postupak hidrolize je primenjen u proizvodnji hidrolizata pivskog tropa koji je kori&scaron;ćen u mlečno-kiselim fermentacijama.<br />Nakon toga je ispitana mlečno-kisela fermentacija sa dva proizvodna mikoorganizma. Kao proizvodni mikroorganizmi u mlečno-kiselim fermentacijama primenjena su dva soja bakterija mlečne kiseline: Lactobacillus fermentum PL-1 i Lactobacillus rhamnosus ATCC 7469. Ispitan je uticaj dodatka različitih koncentracija ekstrakta kvasca (0,5-5,0%) uz korekciju pH vrednosti tokom fermentacije sa dodatkom kalcijum-karbonata. U zavisnosti od udela L-(+)- i D-(-)-mlečne kiseline koje nastaju tokom fermentacije izabran je proizvodni mikroorganizam koji proizvodi vi&scaron;e L-(+)-mlečne kiseline.<br />U daljim ispitivanjima je ispitan uticaj korekcije pH pomoću natrijum-hidroksida kao i dodatak različitih koncentracija ekstrakta kvasca (0,5-5,0%) i redukujućih &scaron;ećera (2,7; 5,4 i 8,1%) u hidrolizatu pivskog tropa na mlečno-kiselu fermentaciju pomoću odabranog soja bakterija mlečne kiseline. Na osnovu dobijenih rezultata izabrana je najbolja koncentracija redukujućih &scaron;ećera i ekstrakta kvasca koji će se koristiti u daljim istraživanjima.<br />Takođe je ispitana i mogućnost zamene skupog ekstrakta kvasca i glukoze sa obnovljivim sirovinama, kao &scaron;to su pivski kvasac, džibra i bistra džibra.<br />Ispitan je uticaj dodatka različitih koncentracija pivskog kvasca (0,5-5,0%), džibre (5-20%) i bistre džibre (5-50%) pre fermentacije kao i dodatak bistre džibre u dolivnoj fermentaciji, na mlečno-kiselu fermentaciju hidrolizata pivskog tropa.<br />Ispitan je i dolivni postupak fermentacije hidrolizata pivskog tropa dodatkom glukoze, glukoze i ekstrakta kvasca i sladovine. Takođe je ispitana mogućnost izvođenja vi&scaron;e uzastopnih fermentacija sa imobilisanim ćelijama odabranog soja bakterija mlečne kiseline u kalcijum-alginatu.<br />Na osnovu eksperimentalnih rezultata zaključujeno je da je dodatak kalcijum-karbonata imao pozitivan uticaj na proizvodnju mlečne kiseline sa L. fermentum i L. rhamnosus. Sa dodatkom kalcijum-karbonata povećali su se utro&scaron;ak redukujućih &scaron;ećera, koncentracija i prinos mlečne kiseline i vijabilnost ćelija L. fermentum i L. rhamnosus. Ekstrakt kvasca i kalcijum-karbonat su imali značajan uticaj na proizvodnju mlečne kiseline sa L. fermentum i L. rhamnosus. U fermentacijama sa L. fermentum najveći prinos ukupne mlečne kiseline (44%) je postignut sa dodatkom 5,0% ekstrakta kvasca i 2,0% kalcijum-karbonata. U fermentacijama sa L. rhamnosus najveći prinos ukupne mlečne kiseline (98%) i L-(+)-mlečne kiseline (96%) je ostvaren u fermentaciji sa dodatkom 2,0% ekstrakta kvasca i 2,0% kalcijum-karbonata. Na osnovu rezultata odlučeno je da se u daljim ispitivanjima mlečno-kisele fermentacije hidrolizata pivskog tropa kao proizvodni mikoorganizam koristi L. rhamnosus.<br />Primenom natrijum-hidroksida za korekciju pH je skratila fermentaciju za 48 sati a ostvareno je i značajno povećanje zapreminske produktivnosti L-(+)-mlečne kiseline (za 200%, povećanje sa 0,21 na 0,63 g/l&middot;h-1). Korekcija pH u svim daljim istraživanjima je vr&scaron;ena sa dodatkom natrijum-hidroksida.<br />U mlečno-kiselim fermentacijama sa različitim početnim koncentracijama redukujućih &scaron;ećera (2,7; 5,4 i 8,1%) i sa dodatkom različitih koncentracija ekstrakta kvasca (0,5-5,0%), najveći prinos L-(+)-mlečne kiseline i zapreminska produktivnost od 91,29% i 1,69 g/l&middot;h-1, kao i vijabilnost ćelija L. rhamnosus od 9,7&middot;109 CFU/ml ostvareni su u fermentaciji sa početniom koncentracijom redukujućih &scaron;ećera od 5,4% i dodatkom 5,0% ekstrakta kvasca.<br />Na osnovu ostvarenih rezultata u istraživanjima sa dodatkom džibre i dodacima tokom fermentacije kao i u fermentacijama sa imobilisanim ćelijama je kori&scaron;ćen hidrolizat pivskog tropa sa početnom koncentracijom redukujućih &scaron;ećera od 5,4%.<br />U mlečno kiseloj fermentaciji sa dodatkom pivskog kvasca najveći prinos L-(+)-mlečne kiseline (89,01%) i zapreminska produktivnost (0,89 g/l&middot;h-1) L-(+)-mlečne kiseline su ostvareni u fermentaciji sa dodatkom 5,0% pivskog kvasca i korekcijom početne koncentracije redukujućih &scaron;ećera na 5,0%. Na osnovu rezultata utvrđeno je da se može izvr&scaron;iti delimična ili potpuna zamena ekstrakta kvasca pivskim kvascem uz značajno smanjenje cene podloge za mlečno-kiselu fermentaciju, bez značajnog smanjenja efikasnosti mlečno-kisele fermentacije.<br />U mlečno-kiseloj fermentaciji sa dodatkom džibre i bistre džibre najveć koncetracija, prinos i zapreminska produktivnost L-(+)-mlečne kiseline od 31,03 g/l, 86,15% i 0,93 g/l&middot;h-1, ostvareni su u fermentaciji sa dodatkom 50% bistre džibre. Najvi&scaron;a koncentracija, prinos i zapreminska produktivnost L-(+)-mlečne kiseline ostvareni u dolivnoj fermentaciji sa dodatkom glukoze i bistre džibre tokom mlečno-kisele fermentacije su iznosili su 48,02 g/l, 87,82% i 0,96 g/l&middot;h-1.<br />U fermentacijama sa dodatkom nutritijenata tokom mlečno-kisele fermentacije najveća vrednost koncetracije, prinosa i zapreminske produktivnosti L-(+)-mlečne kiseline od 116,08 g/l, 93,32% i 2,04 g/L&middot;h-1, su ostvarene u fermentaciji sa dodatkom glukoze i ekstrakta kvasca tokom fermentacije. Na osnovu rezultata utvrđeno je da se dolivni postupak fermentacije može koristiti u cilju povećanja efikasnosti mlečno-kisele fermentacije.<br />Izvr&scaron;ena je imobilizacija ćelija L. rhamnosus u kalcijum-alginatu uz izuzetno visoku vijabilnost (1010 CFU/ml). Imobilisane ćelije L. rhamnosus su uspe&scaron;no kori&scaron;ćene u tri mlečno-kisele fermentacije. Prinos L-(+)-mlečne kiseline i zapreminska produktivnost su u sve tri fermentacije bili izuzetno visoki, pri čemu su najveći prinos L-(+)-mlečne kiseline i zapreminska produktivnost od 95,2% i 1,76 g/l&middot;h-1, ostvareni u drugoj fermentaciji. Upotrebom imobilisanih ćelija L. rhamnosus je osim povećanja prinosa i zapreminske produktivnosti L-(+)-mlečne kiseline skraćena fermentacija za 12 sati u poređenju sa &scaron;aržnim fermentacijama.</p> / <p>Brewers spent grain represents (BSG) about 85% of the total by-products from brewing process and is available at low price during the whole year. Due to its chemical composition BSG has great potential use in biotechnology and production of high-value products. One of very eco-friendly and economical alternative uses of BSG is in production of lactic acid (LA), since in the last few decades the demand for the LA has significantly risen, mostly because of development of biodegradable lactic polymers, which are eco-friendly and nontoxic.<br />Lactic acid is the most important hydrocarboxylic acid with an asymmetrical carbon atom, widely distributed in nature, and it has shown great potential in fields of food, pharmaceutical, textile, leather and chemical industries.<br />The aim of this doctoral thesis was to investigate the application of BSG in lactic acid production. First, the optimization of enzymatic hydrolysis of BSG was conducted, with the goal to achieve high reducing sugar concentrations, as much as possible, that are necessary on LA fermentation. BSG hydrolysis was conducted by usage of commercial enzymes for degradation of starch and cellulose. Effect of pH value, temperature and enzyme dosage on BSG hydrolysis efficiency was investigated. After the best conditions for BSG hydrolysis were determined, the optimized procedure for BSG hydrolysis was used for the production of BSG hydrolysate that will be used in LA fermentations.<br />After optimization of BSG hydrolysis, LA fermentation by two LA producing microorganisms was investigated. The strains investigated were two LA bacteria strains: Lactobacillus fermentum PL-1 and Lactobacillus rhamnosus ATCC 7469. The effect of yeast extract (0.5; 1.0; 2.0; 3.0; 4.0, and 5.0%) addition in BSG hydrolysate, with the correction of pH value during LA fermentation by the addition of calcium-carbonate, on LA fermentation was investigated. Based on the results achieved for L-(+)- and D-(-)-LA ratio the LAB strains that produced more L-(+)-LA was chosen for further research.<br />In further research the effect of pH correction (with addition of NaOH), yeast extract (0.5, 1.0, 2.0, 3.0, 4.0, and 5.0%) addition and reducing sugar concentration (2.7; 5.4 and 8.1%) in BSG hydrolysate on LA fermentation was investigated. Based on the results achieved the best yeast extract and reducing sugars concentrations was determined and used in further analysis or research. Also the possible replacement of expensive yeast extract and glucose with cheap alternatives, like brewer`s spent grain and stillage was investigated. The effect of brewer`s spent grain (0.5; 1.0; 2.0; 3.0; 4.0, and 5.0%), whole stillage (5, 10, 15 i 20%) and thin stillage (5, 10, 15, 20, 30, 40, 50%) addition before fermentation as well as thin stillage addition in fed-batch fermentation in BSG hydrolysate on LA fermentation were investigated.<br />Also fed-batch fermentation procedure (addition of glucose, glucose and yeast extract and wort during fermentation) was investigated. The possible application of cells immobilized in Ca-alginate for LA fermentation of BSG hydrolysate was also investigated.<br />Based on the results it was concluded that BSG can be successfully utilized as a raw material in production of LA, after optimization of hydrolysis and addition of nitrogen source.<br />According to the results of chemical composition before and after optimized hydrolysis 78.6% of total cellulose was hydrolyzed.<br />Addition of calcium-carbonate had positive effect on LA production by L. fermentum i L. rhamnosus. With the addition of calcium-carbonate reducing sugar utilization, LA yield and concentration and cell viability (both L. fermentum i L. rhamnosus) increased. Addition of calcium-carbonate and yeast extract had a positive effect on LA fermentation by L. fermentum and L. rhamnosus. In LA fermentation by L. fermentum the highest LA yield (44%) was achieved with addition of 5.0% of yeast extract and 2.0% of calcium-carbonate. In L. rhamnosus fermentations the highest total LA yield (98%) and L-(+)-LA yield (96%) was reached when 2.0% of yeast extract and 2.0% of calcium-carbonate were added.<br />Based on the results achieved it was concluded that BSG hydrolysate, with the addition of yeast extract, is a good fermentation media for LA fermentation with L. rhamnosus, and it was decided that L. rhamnosus will be used in further research of LA fermentation on BSG hydrolysate.<br />Addition of NaOH instead of calcium-carbonate for the pH correction shortened the fermentation time by 48 h and increased the L-(+)-LA volumetric productivity (by 200%, from 0.21 to 0.63 g/L&middot;h-1). Based on this results pH correction in further experiments was done by addition of NaOH.<br />In LA fermentation with different reducing sugar (2.7, 5.4 and 8.1%) and yeast extract concentrations (0.5-5.0%), the highest L-(+)-LA yield and volumetric productivity of 91.29%, and 1.69 g/L&middot;h-1, respectively, as well as L. rhamnosus cell viability (9.67 log CFU/mL), were achieved with the reducing sugar content of 5.4% and yeast extract content of 5.0%.<br />Based on this results in further experiment with the addition of stillage, in fed-batch fermentation and fermentation with immobilized cell BSG hydrolysate with 5.4% of reducing sugars and 5.0% yeast extract was used.<br />In fermentation with the addition of brewer&rsquo;s spent yeast the highest L-(+)-LA yield (89.01%) and volumetric productivity (0.89 g/L&middot;h-1) were achieved in the fermentation of BSG hydrolysate with 5.0% of reducing sugar and 5.0% of brewer&rsquo;s yeast. Based on the results achieved it was concluded that yeast extract can be partial or complete replaced by brewer&rsquo;s spent yeast with significant decrease of media cost, without the decrease in LA fermentation efficiency.<br />In fermentation with the addition of thin stillage the highest L-(+)-LA concentration, yield, and volumetric productivity of 31.03 g/L, 86.15%, and 0.93 g/L&middot;h-1, respectively, was obtained in fermentation with the addition of 50% of thin stillage. The highest L-(+)-LA concentration, yield, and volumetric productivity achieved in fed-batch fermentation with the addition of glucose and thin stillage during fermentation, were 48,02 g/L, 87,82% i 0,96 g/L&middot;h-1.<br />In fed-batch fermentation the highest L-(+)-LA concentration, yield, and volumetric productivity of 116.08 g/L, 93.32%, and, 2.04 g/L h-1, respectively, were achieved in fermentation with glucose and yeast extract addition during fermentation. The results showerd that fed-batch fermentation could be used to increase L-(+)-LA fermentation efficiency<br />Immobilization of L. rhamnosus cells with high viability (1010 CFU/mL) in Ca-alginate was conducted. Immobilized cells we successfully utilized in three repeated batch fermentation. L-(+)-LA yield and volumetric productivity were very high in all three batch fermentation, with the highest results achieved (95.20% and 1.76 g/L&middot;h-1, respectively) in second fermentation. Application of immobilized L. rhamnosus cells increased L-(+)-LA yield and volumetric productivity and shortened the fermentation time for 12 h in comparison with batch fermentation.</p>