Made available in DSpace on 2017-07-10T19:25:13Z (GMT). No. of bitstreams: 1 henry.pdf: 2217836 bytes, checksum: f01d367fdd6ecd64b7d0ccf6ae52d669 (MD5) Previous issue date: 2012-07-02 / In Brazil, the grain harvest in 2007/08 was about 145 million tons, a record for the domestic agriculture. In 2008 soybeans led to participation in the value of production of temporary and permanent crops with 39.18% which were superseded by the 2009/2010, in which the Paraná state reached the first place in soybean production in Brazil. As possible benefits of diets containing soy we could mention the anticarcinogenic effects, reduction of cholesterol levels, protective effects against obesity and symptoms such as hot flashes during menopause, treatment of coronary heart disease and osteoporosis. According to this economic and health context, this work aimed the development of a hydrosoluble soybean extract fermented with probiotic microorganisms, plus inulin (prebiotic substance) and whey protein concentrate, in order to meet the consumers expectations on health, nutrition and functionality due to their symbiotic nature. Physical and chemical, microbiological, sensory analyzes and probiotic bacteria counting were carried out, considering different types of sugars such as sucrose and glucose. Twelve formulations were prepared from hydrosoluble soybean extract with whey protein concentrate and inulin, by varying the inoculated cultures (Lactobacillus acidophilus, Lactobacillus casei and SAB) and sugar (100% glucose treatment, 100% sucrose treatment, 50% glucose and 50% sucrose treatment, and 0% sugar treatment). The sensory evaluation experiment was completely randomized, applying analysis of variance-ANOVA and Means Test (Tuckey). In the fermentation process the formulation A5 (EHS 10%, 2.5% WPC, 2% inulin, sugar and inoculum SAB) presented the shortest time register, with 5 hours and 53 minutes to achieve the optimum pH. For an optimal therapeutic effect, it is estimated that the probiotic food must contain a number above than 107 CFU / mL, and all formulations reached this viable cell count during 28 days of storage, especially the Treatment A9 with an initial count of 1,20 x1014 CFU / mL. The increased acidity values reached 85 ºD, maintaining the pH value close to 4.31, which means a suitable pH. The results of the microbiological analyzes of fermented beverages were in accordance to the standards established by the Brazilian legislation, ensuring the safety of the samples. The rate of acceptance achieved by sensory evaluation of the beverages was above 70% considering the formulations A2 (EHS 10%, 2.5% WPC, 2% of inulin, 100% sucrose and L. casei inoculum) and A10 (EHS 10% 2.5% WPC, 2% of inulin, and 100% sucrose and L. acidophilus inoculum), which means that these formulations would be well accepted in the consumers market. / A safra brasileira de grãos 2007/08 foi de cerca de 145 milhões de toneladas, um recorde para a agricultura interna. No ano de 2008 a soja liderou a participação no valor da produção de lavouras temporárias e permanentes com 39,18%, participação que foi superada pela safra 2009/2010, na qual o Paraná garantiu o primeiro lugar na produção de soja no Brasil. Como possíveis benefícios de dietas contendo soja podem ser mencionados os efeitos anticarcinogênicos, redução dos níveis de colesterol, efeitos protetores contra a obesidade e sintomas como ondas de calor na menopausa, tratamento de doenças coronarianas e osteoporose. Diante deste contexto econômico e de saúde, almejou-se desenvolver uma bebida à base de extrato hidrossolúvel de soja fermentada com micro-organismos probióticos, acrescida de inulina (substância prebiótica) e proteína concentrada do soro do leite, visando atender às expectativas de consumidores quanto à saúde, nutrição e funcionalidade, devido ao seu caráter simbiótico. Foram realizadas análises físico-químicas, microbiológicas, sensoriais e contagem de bactérias probióticas, considerando-se diferentes tipos de açúcares, como sacarose e glicose. Foram elaboradas doze formulações a partir de extrato hidrossolúvel de soja, adicionado de proteína concentrada de soro de leite e inulina, variando as culturas inoculadas (fermentos lácteos Lactobacillus acidophilus, Lactobacillus casei e SAB) e o açúcar (Tratamento com 100% de glicose, tratamento com 100% de sacarose, tratamento com 50% de glicose e 50% de sacarose e tratamento com 0% de açúcar). O delineamento experimental na avaliação sensorial foi inteiramente casualizado, aplicando-se a análise de variância (ANOVA) e teste de médias (Tuckey). No processo de fermentação a formulação contendo 10% EHS, 2,5% WPC, 2% de inulina, sem açúcar e o inóculo SAB sobressaiu-se com o menor tempo, totalizando 5 horas e 53 minutos para alcançar o pH ótimo. Para obter um efeito terapêutico ótimo, estima-se que o alimento probiótico deva conter um número maior que 107 UFC/mL, sendo que todas as formulações elaboradas alcançaram este propósito durante os 28 dias de armazenamento, tendo como destaque a formulação A9 com uma contagem inicial de 1,20 x1014 UFC/mL. Os valores de acidez aumentaram até aproximadamente 85º D, mantendo o valor de pH próximo a 4,31, o que significa um pH adequado. Os resultados das análises microbiológicas das bebidas fermentadas apresentaram-se em conformidade com os padrões estabelecidos pela legislação brasileira, assegurando a inocuidade das amostras. O índice de aceitabilidade obtido pela avaliação sensorial das bebidas foi superior a 70% para as formulações A2 (10% EHS, 2,5% WPC, 2% de inulina, 100% sacarose e o inóculo L. casei ) e A10 (10% EHS, 2,5%WPC,2% de inulina, 100%sacarose e o inóculo L. acidófilos), o que significa que estas formulações seriam bem aceitas no mercado consumidor.

Made available in DSpace on 2017-05-12T14:48:38Z (GMT). No. of bitstreams: 1 henry.pdf: 2217836 bytes, checksum: f01d367fdd6ecd64b7d0ccf6ae52d669 (MD5) Previous issue date: 2012-07-02 / In Brazil, the grain harvest in 2007/08 was about 145 million tons, a record for the domestic agriculture. In 2008 soybeans led to participation in the value of production of temporary and permanent crops with 39.18% which were superseded by the 2009/2010, in which the Paraná state reached the first place in soybean production in Brazil. As possible benefits of diets containing soy we could mention the anticarcinogenic effects, reduction of cholesterol levels, protective effects against obesity and symptoms such as hot flashes during menopause, treatment of coronary heart disease and osteoporosis. According to this economic and health context, this work aimed the development of a hydrosoluble soybean extract fermented with probiotic microorganisms, plus inulin (prebiotic substance) and whey protein concentrate, in order to meet the consumers expectations on health, nutrition and functionality due to their symbiotic nature. Physical and chemical, microbiological, sensory analyzes and probiotic bacteria counting were carried out, considering different types of sugars such as sucrose and glucose. Twelve formulations were prepared from hydrosoluble soybean extract with whey protein concentrate and inulin, by varying the inoculated cultures (Lactobacillus acidophilus, Lactobacillus casei and SAB) and sugar (100% glucose treatment, 100% sucrose treatment, 50% glucose and 50% sucrose treatment, and 0% sugar treatment). The sensory evaluation experiment was completely randomized, applying analysis of variance-ANOVA and Means Test (Tuckey). In the fermentation process the formulation A5 (EHS 10%, 2.5% WPC, 2% inulin, sugar and inoculum SAB) presented the shortest time register, with 5 hours and 53 minutes to achieve the optimum pH. For an optimal therapeutic effect, it is estimated that the probiotic food must contain a number above than 107 CFU / mL, and all formulations reached this viable cell count during 28 days of storage, especially the Treatment A9 with an initial count of 1,20 x1014 CFU / mL. The increased acidity values reached 85 ºD, maintaining the pH value close to 4.31, which means a suitable pH. The results of the microbiological analyzes of fermented beverages were in accordance to the standards established by the Brazilian legislation, ensuring the safety of the samples. The rate of acceptance achieved by sensory evaluation of the beverages was above 70% considering the formulations A2 (EHS 10%, 2.5% WPC, 2% of inulin, 100% sucrose and L. casei inoculum) and A10 (EHS 10% 2.5% WPC, 2% of inulin, and 100% sucrose and L. acidophilus inoculum), which means that these formulations would be well accepted in the consumers market. / A safra brasileira de grãos 2007/08 foi de cerca de 145 milhões de toneladas, um recorde para a agricultura interna. No ano de 2008 a soja liderou a participação no valor da produção de lavouras temporárias e permanentes com 39,18%, participação que foi superada pela safra 2009/2010, na qual o Paraná garantiu o primeiro lugar na produção de soja no Brasil. Como possíveis benefícios de dietas contendo soja podem ser mencionados os efeitos anticarcinogênicos, redução dos níveis de colesterol, efeitos protetores contra a obesidade e sintomas como ondas de calor na menopausa, tratamento de doenças coronarianas e osteoporose. Diante deste contexto econômico e de saúde, almejou-se desenvolver uma bebida à base de extrato hidrossolúvel de soja fermentada com micro-organismos probióticos, acrescida de inulina (substância prebiótica) e proteína concentrada do soro do leite, visando atender às expectativas de consumidores quanto à saúde, nutrição e funcionalidade, devido ao seu caráter simbiótico. Foram realizadas análises físico-químicas, microbiológicas, sensoriais e contagem de bactérias probióticas, considerando-se diferentes tipos de açúcares, como sacarose e glicose. Foram elaboradas doze formulações a partir de extrato hidrossolúvel de soja, adicionado de proteína concentrada de soro de leite e inulina, variando as culturas inoculadas (fermentos lácteos Lactobacillus acidophilus, Lactobacillus casei e SAB) e o açúcar (Tratamento com 100% de glicose, tratamento com 100% de sacarose, tratamento com 50% de glicose e 50% de sacarose e tratamento com 0% de açúcar). O delineamento experimental na avaliação sensorial foi inteiramente casualizado, aplicando-se a análise de variância (ANOVA) e teste de médias (Tuckey). No processo de fermentação a formulação contendo 10% EHS, 2,5% WPC, 2% de inulina, sem açúcar e o inóculo SAB sobressaiu-se com o menor tempo, totalizando 5 horas e 53 minutos para alcançar o pH ótimo. Para obter um efeito terapêutico ótimo, estima-se que o alimento probiótico deva conter um número maior que 107 UFC/mL, sendo que todas as formulações elaboradas alcançaram este propósito durante os 28 dias de armazenamento, tendo como destaque a formulação A9 com uma contagem inicial de 1,20 x1014 UFC/mL. Os valores de acidez aumentaram até aproximadamente 85º D, mantendo o valor de pH próximo a 4,31, o que significa um pH adequado. Os resultados das análises microbiológicas das bebidas fermentadas apresentaram-se em conformidade com os padrões estabelecidos pela legislação brasileira, assegurando a inocuidade das amostras. O índice de aceitabilidade obtido pela avaliação sensorial das bebidas foi superior a 70% para as formulações A2 (10% EHS, 2,5% WPC, 2% de inulina, 100% sacarose e o inóculo L. casei ) e A10 (10% EHS, 2,5%WPC,2% de inulina, 100%sacarose e o inóculo L. acidófilos), o que significa que estas formulações seriam bem aceitas no mercado consumidor.

The effect of inclusion rate of pea meal (PM), pea protein concentrate (PPC), soybean meal (SBM), soy protein concentrate (SPC), canola meal (CM) and canola protein concentrate (CPC) in salmonid diets was determined through six corresponding meta-analyses of all data available in the literature for these six feed ingredients, which was followed by weighted regression analysis. Increasing dietary inclusion levels of SBM, SPC, CM and CPC reduced specific growth rate (SGR). Regression analysis determined all of these relationships to be linear declines in SGR (P < 0.05). Inclusion levels of PM or PPC did not influence salmonid SGR (P > 0.05). These results showed that the influence plant proteins have on salmonid SGR is dependent on ingredient type and inclusion level. PM, PPC, SBM, SPC, CM an aqueous-extracted CPC and a high phytate CPC (PCPC) were analyzed for chemical nutrient (proximate, amino acid and phosphorus analysis) and antinutrient composition and total tract digestibility (two separate digestibility trials) in rainbow trout (Oncorhynchus mykiss). The digestibility of proximate chemical components and amino acids were significantly higher for the soy products than the pea products. These digestibilities were also significantly higher in protein concentrates than in plant meals. Dry matter and gross energy digestibility was higher in CPC than in CM (P < 0.05). Phosphorus digestibility was higher in CPC-fed fish than in CM-fed fish (P < 0.05), which is likely due to the fact that CPC did not contain phytic acid. Six consecutive growth studies (one trial per test ingredient) were conducted over a 361-day period to determine the effects of feeding increasing inclusion rates of PM, PPC, SBM, SPC, CM and CPC on the growth performance of rainbow trout. Diets were formulated based on the digestible nutrient content of all ingredients as determined in the previously conducted digestibility trials, to contain 0, 75, 150, 225 or 300 g/kg of each test ingredient. All diets were nutritionally equal and contained 17.6 MJ/kg digestible energy, 386.2 g/kg digestible crude protein and were balanced for digestible essential amino acids to meet or exceed the requirements of rainbow trout. Linear and quadratic analysis was conducted on the experimental data. There were no significant regressions resulting from feeding PM, SPC or CPC at 0-300 g/kg for average daily feed intake (ADFI), specific growth rate (SGR), feed conversion ratio (FCR) or protein efficiency ratio (PER). A positive relationship was associated between PPC inclusion and ADFI (P < 0.05). There was a significantly negative quadratic equation associated with the inclusion level of SBM on SGR and FCR and significantly negative linear and quadratic equations for PER. There were significantly negative linear relationships between the inclusion rate of CM and the SGR, FCR and PER of rainbow trout (P < 0.05). Growth trial results suggest at inclusion levels up to 300 g/kg, PM, PPC, SPC and CPC are feasible plant-based fish meal replacements with predictable growth effects, provided the nutritional constraints set in this experiment are followed. The results of these growth experiments were further analyzed using structural equation modeling to determine the relationship between ANF in the six ingredients and ADFI and SGR, which were transformed (tSGR and tADFI, respectively) to enable comparisons between experiments. All possible models between ingredient ANF (starch, phytic acid, glucosinolates, tannins, isoflavones, total NSP, soluble NSP, insoluble NSP and saponins) and ADFI and SGR were calculated. The model with the highest likelihood, as determined by the Akaike Information Criteria0, contained 29 parameters and six degrees of freedom. tADFI positively influenced tSGR. Glucosinolates, saponins, and phytates had a significantly negative impact on tADFI, whereas tannins had a significantly positive impact. The presence of saponins in the diet resulted in a decrease in tSGR. This structural equation model had significant correlations between all ANF, with the exception of phytates and saponins. Future applications of this work will be to develop a nutritional model for optimal inclusion of plant-based feed ingredients in rainbow trout diets, based on their ANF content, which may improve the accuracy of diet formulation and growth prediction.

Milk is provided to new borns as their first food source and it contains essential nutrients, vitamins and other beneficial components, such as enzymes and antibodies that are required for rapid growth and development of the new born and for sustained growth over time. Milk contains two main types of proteins; casein proteins and whey proteins. Although casein proteins account for up to 80% of the proteins found in bovine milk, it is the whey protein that has become of high interest because of its bioactive content. Whey, a very watery mixture of lactose, proteins, minerals and trace amounts of fat, is formed from milk when the milk is coagulated and/or the casein proteins are removed from the milk. Bovine whey protein, including both the acidic and basic fractions (low and high isoelectric point, respectively), has previously been studied in vitro (cell based) and in vivo (using rats) for its impact on bone to determine if it can help improve bone mineral density and help reduce the risk of developing bone diseases, such as osteoporosis. Bone is constantly undergoing a remodelling process of being dissolved and reformed and the two main cell types responsible for this bone remodelling process are mature osteoclasts, which dissolve (resorb) bone, and osteoblasts, which reform the bone. Prior work has shown that acidic protein fractions derived from different sources of whey protein concentrate (WPC) have both in vivo and in vitro activity on bone, particularly anti-resorptive properties. However, the component(s) which confer activity have not yet been identified. In this thesis, work was undertaken to better understand the analytical composition of three types of WPC (cheese, mineral acid and lactic acid) and their associated acidic protein fractions and relate this to bone activity in the hope of identifying where the activity lies. Bone activity was assessed using in vitro screening with osteoblast cells (MC3T3-E1) and osteoclast cells (RAW 264.7). Comparison of the cell-based bone activity of the parent WPCs and corresponding acidic fractions indicated that the acidic fractions derived from both mineral acid and lactic WPC were superior in their ability to inhibit osteoclast development. Although compositional data was complex and definitive correlations with both bone bioactivities could not be made, it appeared that elements common to both the acidic fractions were a higher proportion of GLYCAM-1 and bone sialoprotein-1 (osteopontin). Further studies to more closely investigate the bone bioactivity of the acidic fractions are warranted.

Milk is provided to new borns as their first food source and it contains essential nutrients, vitamins and other beneficial components, such as enzymes and antibodies that are required for rapid growth and development of the new born and for sustained growth over time. Milk contains two main types of proteins; casein proteins and whey proteins. Although casein proteins account for up to 80% of the proteins found in bovine milk, it is the whey protein that has become of high interest because of its bioactive content. Whey, a very watery mixture of lactose, proteins, minerals and trace amounts of fat, is formed from milk when the milk is coagulated and/or the casein proteins are removed from the milk. Bovine whey protein, including both the acidic and basic fractions (low and high isoelectric point, respectively), has previously been studied in vitro (cell based) and in vivo (using rats) for its impact on bone to determine if it can help improve bone mineral density and help reduce the risk of developing bone diseases, such as osteoporosis. Bone is constantly undergoing a remodelling process of being dissolved and reformed and the two main cell types responsible for this bone remodelling process are mature osteoclasts, which dissolve (resorb) bone, and osteoblasts, which reform the bone. Prior work has shown that acidic protein fractions derived from different sources of whey protein concentrate (WPC) have both in vivo and in vitro activity on bone, particularly anti-resorptive properties. However, the component(s) which confer activity have not yet been identified. In this thesis, work was undertaken to better understand the analytical composition of three types of WPC (cheese, mineral acid and lactic acid) and their associated acidic protein fractions and relate this to bone activity in the hope of identifying where the activity lies. Bone activity was assessed using in vitro screening with osteoblast cells (MC3T3-E1) and osteoclast cells (RAW 264.7). Comparison of the cell-based bone activity of the parent WPCs and corresponding acidic fractions indicated that the acidic fractions derived from both mineral acid and lactic WPC were superior in their ability to inhibit osteoclast development. Although compositional data was complex and definitive correlations with both bone bioactivities could not be made, it appeared that elements common to both the acidic fractions were a higher proportion of GLYCAM-1 and bone sialoprotein-1 (osteopontin). Further studies to more closely investigate the bone bioactivity of the acidic fractions are warranted.

Milk is provided to new borns as their first food source and it contains essential nutrients, vitamins and other beneficial components, such as enzymes and antibodies that are required for rapid growth and development of the new born and for sustained growth over time. Milk contains two main types of proteins; casein proteins and whey proteins. Although casein proteins account for up to 80% of the proteins found in bovine milk, it is the whey protein that has become of high interest because of its bioactive content. Whey, a very watery mixture of lactose, proteins, minerals and trace amounts of fat, is formed from milk when the milk is coagulated and/or the casein proteins are removed from the milk. Bovine whey protein, including both the acidic and basic fractions (low and high isoelectric point, respectively), has previously been studied in vitro (cell based) and in vivo (using rats) for its impact on bone to determine if it can help improve bone mineral density and help reduce the risk of developing bone diseases, such as osteoporosis. Bone is constantly undergoing a remodelling process of being dissolved and reformed and the two main cell types responsible for this bone remodelling process are mature osteoclasts, which dissolve (resorb) bone, and osteoblasts, which reform the bone. Prior work has shown that acidic protein fractions derived from different sources of whey protein concentrate (WPC) have both in vivo and in vitro activity on bone, particularly anti-resorptive properties. However, the component(s) which confer activity have not yet been identified. In this thesis, work was undertaken to better understand the analytical composition of three types of WPC (cheese, mineral acid and lactic acid) and their associated acidic protein fractions and relate this to bone activity in the hope of identifying where the activity lies. Bone activity was assessed using in vitro screening with osteoblast cells (MC3T3-E1) and osteoclast cells (RAW 264.7). Comparison of the cell-based bone activity of the parent WPCs and corresponding acidic fractions indicated that the acidic fractions derived from both mineral acid and lactic WPC were superior in their ability to inhibit osteoclast development. Although compositional data was complex and definitive correlations with both bone bioactivities could not be made, it appeared that elements common to both the acidic fractions were a higher proportion of GLYCAM-1 and bone sialoprotein-1 (osteopontin). Further studies to more closely investigate the bone bioactivity of the acidic fractions are warranted.

Milk is provided to new borns as their first food source and it contains essential nutrients, vitamins and other beneficial components, such as enzymes and antibodies that are required for rapid growth and development of the new born and for sustained growth over time. Milk contains two main types of proteins; casein proteins and whey proteins. Although casein proteins account for up to 80% of the proteins found in bovine milk, it is the whey protein that has become of high interest because of its bioactive content. Whey, a very watery mixture of lactose, proteins, minerals and trace amounts of fat, is formed from milk when the milk is coagulated and/or the casein proteins are removed from the milk. Bovine whey protein, including both the acidic and basic fractions (low and high isoelectric point, respectively), has previously been studied in vitro (cell based) and in vivo (using rats) for its impact on bone to determine if it can help improve bone mineral density and help reduce the risk of developing bone diseases, such as osteoporosis. Bone is constantly undergoing a remodelling process of being dissolved and reformed and the two main cell types responsible for this bone remodelling process are mature osteoclasts, which dissolve (resorb) bone, and osteoblasts, which reform the bone. Prior work has shown that acidic protein fractions derived from different sources of whey protein concentrate (WPC) have both in vivo and in vitro activity on bone, particularly anti-resorptive properties. However, the component(s) which confer activity have not yet been identified. In this thesis, work was undertaken to better understand the analytical composition of three types of WPC (cheese, mineral acid and lactic acid) and their associated acidic protein fractions and relate this to bone activity in the hope of identifying where the activity lies. Bone activity was assessed using in vitro screening with osteoblast cells (MC3T3-E1) and osteoclast cells (RAW 264.7). Comparison of the cell-based bone activity of the parent WPCs and corresponding acidic fractions indicated that the acidic fractions derived from both mineral acid and lactic WPC were superior in their ability to inhibit osteoclast development. Although compositional data was complex and definitive correlations with both bone bioactivities could not be made, it appeared that elements common to both the acidic fractions were a higher proportion of GLYCAM-1 and bone sialoprotein-1 (osteopontin). Further studies to more closely investigate the bone bioactivity of the acidic fractions are warranted.

Milk is provided to new borns as their first food source and it contains essential nutrients, vitamins and other beneficial components, such as enzymes and antibodies that are required for rapid growth and development of the new born and for sustained growth over time. Milk contains two main types of proteins; casein proteins and whey proteins. Although casein proteins account for up to 80% of the proteins found in bovine milk, it is the whey protein that has become of high interest because of its bioactive content. Whey, a very watery mixture of lactose, proteins, minerals and trace amounts of fat, is formed from milk when the milk is coagulated and/or the casein proteins are removed from the milk. Bovine whey protein, including both the acidic and basic fractions (low and high isoelectric point, respectively), has previously been studied in vitro (cell based) and in vivo (using rats) for its impact on bone to determine if it can help improve bone mineral density and help reduce the risk of developing bone diseases, such as osteoporosis. Bone is constantly undergoing a remodelling process of being dissolved and reformed and the two main cell types responsible for this bone remodelling process are mature osteoclasts, which dissolve (resorb) bone, and osteoblasts, which reform the bone. Prior work has shown that acidic protein fractions derived from different sources of whey protein concentrate (WPC) have both in vivo and in vitro activity on bone, particularly anti-resorptive properties. However, the component(s) which confer activity have not yet been identified. In this thesis, work was undertaken to better understand the analytical composition of three types of WPC (cheese, mineral acid and lactic acid) and their associated acidic protein fractions and relate this to bone activity in the hope of identifying where the activity lies. Bone activity was assessed using in vitro screening with osteoblast cells (MC3T3-E1) and osteoclast cells (RAW 264.7). Comparison of the cell-based bone activity of the parent WPCs and corresponding acidic fractions indicated that the acidic fractions derived from both mineral acid and lactic WPC were superior in their ability to inhibit osteoclast development. Although compositional data was complex and definitive correlations with both bone bioactivities could not be made, it appeared that elements common to both the acidic fractions were a higher proportion of GLYCAM-1 and bone sialoprotein-1 (osteopontin). Further studies to more closely investigate the bone bioactivity of the acidic fractions are warranted.

Milk is provided to new borns as their first food source and it contains essential nutrients, vitamins and other beneficial components, such as enzymes and antibodies that are required for rapid growth and development of the new born and for sustained growth over time. Milk contains two main types of proteins; casein proteins and whey proteins. Although casein proteins account for up to 80% of the proteins found in bovine milk, it is the whey protein that has become of high interest because of its bioactive content. Whey, a very watery mixture of lactose, proteins, minerals and trace amounts of fat, is formed from milk when the milk is coagulated and/or the casein proteins are removed from the milk. Bovine whey protein, including both the acidic and basic fractions (low and high isoelectric point, respectively), has previously been studied in vitro (cell based) and in vivo (using rats) for its impact on bone to determine if it can help improve bone mineral density and help reduce the risk of developing bone diseases, such as osteoporosis. Bone is constantly undergoing a remodelling process of being dissolved and reformed and the two main cell types responsible for this bone remodelling process are mature osteoclasts, which dissolve (resorb) bone, and osteoblasts, which reform the bone. Prior work has shown that acidic protein fractions derived from different sources of whey protein concentrate (WPC) have both in vivo and in vitro activity on bone, particularly anti-resorptive properties. However, the component(s) which confer activity have not yet been identified. In this thesis, work was undertaken to better understand the analytical composition of three types of WPC (cheese, mineral acid and lactic acid) and their associated acidic protein fractions and relate this to bone activity in the hope of identifying where the activity lies. Bone activity was assessed using in vitro screening with osteoblast cells (MC3T3-E1) and osteoclast cells (RAW 264.7). Comparison of the cell-based bone activity of the parent WPCs and corresponding acidic fractions indicated that the acidic fractions derived from both mineral acid and lactic WPC were superior in their ability to inhibit osteoclast development. Although compositional data was complex and definitive correlations with both bone bioactivities could not be made, it appeared that elements common to both the acidic fractions were a higher proportion of GLYCAM-1 and bone sialoprotein-1 (osteopontin). Further studies to more closely investigate the bone bioactivity of the acidic fractions are warranted.

Milk is provided to new borns as their first food source and it contains essential nutrients, vitamins and other beneficial components, such as enzymes and antibodies that are required for rapid growth and development of the new born and for sustained growth over time. Milk contains two main types of proteins; casein proteins and whey proteins. Although casein proteins account for up to 80% of the proteins found in bovine milk, it is the whey protein that has become of high interest because of its bioactive content. Whey, a very watery mixture of lactose, proteins, minerals and trace amounts of fat, is formed from milk when the milk is coagulated and/or the casein proteins are removed from the milk. Bovine whey protein, including both the acidic and basic fractions (low and high isoelectric point, respectively), has previously been studied in vitro (cell based) and in vivo (using rats) for its impact on bone to determine if it can help improve bone mineral density and help reduce the risk of developing bone diseases, such as osteoporosis. Bone is constantly undergoing a remodelling process of being dissolved and reformed and the two main cell types responsible for this bone remodelling process are mature osteoclasts, which dissolve (resorb) bone, and osteoblasts, which reform the bone. Prior work has shown that acidic protein fractions derived from different sources of whey protein concentrate (WPC) have both in vivo and in vitro activity on bone, particularly anti-resorptive properties. However, the component(s) which confer activity have not yet been identified. In this thesis, work was undertaken to better understand the analytical composition of three types of WPC (cheese, mineral acid and lactic acid) and their associated acidic protein fractions and relate this to bone activity in the hope of identifying where the activity lies. Bone activity was assessed using in vitro screening with osteoblast cells (MC3T3-E1) and osteoclast cells (RAW 264.7). Comparison of the cell-based bone activity of the parent WPCs and corresponding acidic fractions indicated that the acidic fractions derived from both mineral acid and lactic WPC were superior in their ability to inhibit osteoclast development. Although compositional data was complex and definitive correlations with both bone bioactivities could not be made, it appeared that elements common to both the acidic fractions were a higher proportion of GLYCAM-1 and bone sialoprotein-1 (osteopontin). Further studies to more closely investigate the bone bioactivity of the acidic fractions are warranted.