Nutritional and Phytochemical Content of High-Protein Crops

Peer reviewed

Infant nutrition and the gut microbiota

Establishment of the intestinal microbiota commences at birth and this colonisation is influenced by a number of factors including mode of delivery, gestational age, mode of feeding, environmental factors and host genetics. As this initial establishment may well influence the health of an individual later in life, it is imperative to understand this process. Therefore, this thesis set out to investigate how early infant nutrition influences the development of a healthy gut microbiota. As part of the INFANTMET project, the intestinal microbiota of 199 breastfed infants was investigated using both culture-dependent and culture-independent approaches. This study revealed that delivery mode and gestational age had a significant impact on early microbial communities. In order to understand host genotype-microbiota interactions, the gut microbiota composition of dichorionic triplets was also investigated. The results suggested that initially host genetics play a significant role in the composition of an individual’s gut microbiota, but by month 12 environmental factors are the major determinant. To investigate the origin of hydrogen sulphide in a case of nondrug- induced sulfhemoglobinemia in a preterm infant, the gut microbiota composition was determined. This analysis revealed the presence of Morganella morganii, a producer of hydrogen sulphide and hemolysins, at a relative abundance 38%, which was not detected in control infants. Following on from this, the negative and short term consequences of intrapartum antibiotic prophylaxis exposure on the early infant intestinal microbiota composition were demonstrated, particularly in breast-fed infants, which are recovered by day 30. Finally, the composition of the breast milk microbiota over the first three months of life was characterised. A core of 12 genera were identified amongst women and the remainder comprised some 195 genera which were individual specific and subject to variations over time. The results presented in this thesis have demonstrated that the development of the infant gut microbiota is complex and highly individual. Clear alterations in the intestinal microbiota establishment process in C-section delivered, preterm and antibiotic exposed infants were shown. Taken together, long-term health benefits for infants, particularly those vulnerable groups, may be conferred through the design of probiotic and prebiotic food ingredients and supplements.

Effects of early nutrition on the wnt and bone morphogenic protein pathways in the developing intestine

Infant formula is consumed by the majority of infants in the United States for at least part of the first year of life. Infant formula lacks many of the bioactive compounds that are naturally occurring in breast milk. Because of this, there has been an increased interest by the companies that manufacture infant formula to include additives that would potentially allow formula to more closely mimic breast milk activity. One such ingredient currently being added to infant formula is prebiotics. Prebiotics are non-digestible food ingredients that beneficially affect the host by selectively stimulating the growth of specific healthful bacteria in the colon. It is speculated that prebiotics replicate the activity of breast milk oligosaccharides, which through the production of butyrate by intestinal microbiota, may interact with the Wnt/BMP pathways. The Wnt/BMP pathways regulate intestinal stem cells, which determine the growth, development and maintenance of the intestine. Therefore, the objective of this study was to explore the effects that the addition of prebiotics to formula have on the regulation of the Wnt/BMP pathways when fed to neonatal piglets, a model commonly used in the study of infant nutrition. Piglets (n=5) were randomized into sow-reared (SR), fed control formula (F), or fed formula with added prebiotics (F+P). Fructooligosaccharides (FOS) (2 g/L) and polydextrose (PDX) (2 g/L) were chosen as the prebiotics for this study, because this combination had been less studied than other combinations. Ileum and ascending colon were collected at 7 and 14 days-of-age. Dry matter content, pH, and short chain fatty acid (SCFA) content was measured. The mRNA expression of β-catenin, sFRP3, sFRP4, frizzled 6, DKK1 (Wnt pathway), gremlin (BMP pathway), TNF-a, HNF-4α and osteopontin (OPN) was measured by RT-qPCR. Piglets fed the F+P diet had greater acetate concentration and lower pH in the ileum at day 14 and in the colon at day 7 and day 14 than F piglets. Butyrate concentrations were highest in SR with F+P not differing from F in ileum at day 14 and colon at day 7 and day 14. Effects of age were seen in all genes, with the exception of OPN, sFRP-3 and sFRP-4. On day 7, no effect of diet was observed in the ileum, however, mRNA expression of DKK1 and frizzled 6 were greater in F+P than SR (p≤0.05). On day 14, gremlin expression was lower and OPN was greater in the ileum of SR piglets compared to F and F+P. Also on day 14, HNF-4α mRNA expression was greater in both ileum and colon of F+P piglets and sFRP3 mRNA expression was greater in the colon than F or SR . In summary, differences were observed between gene expression of F+P and SR piglet intestines, but the supplementation of 2 g/L scFOS and 2 g/L PDX to formula did not shift expression of genes in the Wnt/BMP pathways to be more similar to SR than F. As the Wnt/BMP pathway is known to exist in a gradient along the crypt-villus axis, with Wnt expression dominating in the crypt region and BMP expression dominating in the villi, it was possible that pooling whole tissue reduced our ability to detect treatment effects that would be concentrated in either region. A method was therefore developed to remove intestinal epithelial cells along the villus-to-crypt axis. Twenty-five-day-old F and SR piglets were euthanized and ileal tissue was collected and placed in a dissociation buffer in a shaking water bath. Exfoliated cells were removed at increasing time points from 5 to 100 minutes in order to remove cells along the villus-to-crypt axis. After the final incubation, remaining mucosal tissue was removed using a sterile glass microscope slide and pooled with the final exfoliated cell isolation. After each cell collection, a section of tissue was fixed in formalin for histomorphological examination. Expression of genes in the Wnt/BMP pathways, along with crypt marker genes (CDK5 and v-myb), were measured in both whole ileal tissue, pooled epithelial cells, and separate epithelial cell isolations from the same piglet. The expression of β-catenin, HNF-4α, TNF-α, TGF-β and the crypt marker v-myb matched the expected villus-to-crypt pattern in cells collected after 10 (incubation 1), 30 (incubation 2) and 60 (incubation 3) minutes. However, expression of expression in cells collected after 100 minutes (incubation 4) was variable, which may be due to the fact that crypt cells were not efficiently removed and the presence of unwanted non-epithelial tissue. Gremlin, OPN, DKK1, sFRP3 and sFRP4 expression was not statistically different along the villus-to-crypt axis. Frizzled 6 and CDK5 did not express as we had predicted, with expression highest towards the villi. In summary, the epithelial cell collection method used was not entirely successful. While much of the gene data suggests that cells were removed along the villus-to-crypt axis through the first three incubations, the last incubation, which involved scraping the tissue, removed non-epithelial components of the mucosa, while leaving the crypts intact. In conclusion, the addition of 2 g/L PDX and 2 g/L scFOS did not cause gene expression of the Wnt/BMP pathways to mirror either F or SR expression. New isolation methods to extract cells along the crypt-villus axis should be considered, including the use of a laser capture microdissection. While this combination of prebiotics did not yield the intended effects, future research should be done on other combinations, such as the inclusion of galactooligosaccharides (GOS), which is commonly added to food products including infant formula.

Utilização de bagaço de maçã como ingrediente alimentar em iogurte com potencial efeito benéfico para a saúde

O desenvolvimento de alimentos funcionais evoluiu consideravelmente ao longo dos anos e a capacidade tecnológica para produzir um alimento com compostos fisiologicamente ativos tem crescido significativamente. O presente trabalho teve como objetivo criar um novo alimento, utilizando um subproduto proveniente da indústria agroalimentar. O alimento seleccionado foi o iogurte natural, o qual foi enriquecido com um extrato de bagaço de maçã. O bagaço de maçã contém compostos fenólicos e fibra, mostrando atividade antioxidante significativa e por conseguinte apresenta um potencial efeito positivo na saúde. Avaliaram-se características químicas do bagaço de maçã e das farinhas obtidas a partir deste subproduto, designadamente: acidez, teor em açúcares totais e redutores, cinza, matéria gorda, humidade, proteína bruta, fibra dietética insolúvel e solúvel. O extrato aquoso obtido a partir do subproduto foi também caracterizado quanto ao seu conteúdo fenólico e atividade antioxidante. O iogurte produzido com incorporação do extrato de bagaço de maçã foi estudado do ponto de vista de parâmetros químicos tais como acidez, açúcares totais, cinza, humidade, proteína bruta, pH e fibra bruta, conteúdo em fenólicos totais e atividade antioxidante. O subproduto, os extratos e o iogurte foram também avaliados quanto à sua carga microbiológica. Na caracterização química do bagaço foram obtidos os seguintes valores, expressos em base seca: 2,0±0,01% de acidez (expressa em equivalentes de ácido málico); 15,96±1,53% de açúcares totais; 13,35±1,91% de açúcares redutores; 1,88±0,07% de cinza, 2,49±0,5% de matéria gorda, 81,17±1,98% de humidade e 5,01±0,01% de proteína bruta. Quanto ao seu conteúdo em fibra dietética, o bagaço contém na sua composição 65,80% de fibra dietética insolúvel e 4,90% de fibra dietética solúvel. A farinha obtida após secagem a 60 °C do bagaço de maçã apresentou, na base seca, 1,9±0,04% de acidez, 10,57±1,31% de açúcares totais; 8,50±1,00% de açúcares redutores; 2,22±0,04%de cinza, 4,73±0,11% de matéria gorda, 6,34±0,62% de humidade e 5,40±0,26% de proteína. A atividade antioxidante do extrato aquoso, (AQ_6X10) obtido do bagaço de maçã, utilizado para incorporação no iogurte, determinada através do método ABTS foi de 5,00±1,28µmol TE/g amostra e apresentou um teor em compostos fenólicos de 221,42±0,734 mg EAG/ 100g de extrato, na base seca. Ao nível microbiológico o extrato revelou parâmetros aceitáveis, de acordo com a tabela 13 (anexos 3), para utilização como ingrediente alimentar. Os iogurtes produzidos foram analisados quimicamente. O iogurte, com extrato aquoso de bagaço de maçã incorporado, apresentou 89,64±0,00% de humidade, 0,93±0,00% de acidez (expressa em equivalentes de ácido láctico); 6,42±0,26% de açúcares totais; 0,74%±0,00% de cinza, 3,83±0,00% de proteína bruta e 0,2±0,00% de fibra bruta. O seu conteúdo em compostos fenólicos foi de 12,41±1,69mg EAG/ 100g de iogurte, e a sua atividade antioxidante foi 54,84±4,40 µmol TE/g iogurte. Avaliou-se ainda o iogurte no que diz respeito ao crescimento de bactérias lácticas e constatou-se, por comparação com o iogurte de controlo, que estas se desenvolveram normalmente ao longo do processo de fabrico. A análise microbiológica revelou ainda que o iogurte é seguro do ponto de vista alimentar de acordo com a Tabela 15 (anexos 3). A análise sensorial realizada aos iogurtes demonstrou que o iogurte fortificado apresentou uma aceitação muito boa pelo painel de provadores. O presente estudo demonstrou que o bagaço de maçã, um subproduto das indústrias agroalimentares, é seguro do ponto de vista microbiológico, podendo ser utilizado na preparação de ingredientes alimentares para serem incorporados, por exemplo em iogurte, conferindo-lhe características que se destacam pelo maior teor em fibra e atividade antioxidante em relação ao iogurte não enriquecido, e atributos sensoriais apelativos em termos de textura e sabor.

Probiotics, Prebiotics and Synbiotics

Probiotics are living microorganisms which, when ingested in certain amounts, have a positive impact on human health, mainly due to their roles in improving the balance of the intestinal microflora. On the other hand, the prebiotic are food ingredients that may also have a positive impact in the improvement of the intestinal flora. These components, which fall into the category of fibers, are not digested in the upper gastrointestinal tract, and therefore reach the colon where they stimulate the growth and/or the activity of some types of bacteria. The term synbiotic is used for products that contain both probiotics and prebiotics, thus taking advantage of both the addition of beneficial bacteria and the encouragement of the growth of resident beneficial bacteria. The present chapter aims to review the scientific literature related to prebiotics, probiotics and synbiotics, including their identification, properties and health benefits.

Chemistry and food preparation, a laboratory manual for students of home economics; a series of experiments in elementary chemistry and experimental cookery designed for a scientific approach to the study of food preparation. Basic recipes and methods of combining ingredients are presented, together with variations that suggest the wide applicability of the fundamental principles.

Bibliography: p. 1

The influence of the amount of sunscreen applied and its sun protection factor (SPF): evaluation of two sunscreens including the same ingredients at different concentrations

Background To estimate labeled sun protection factor (SPF) for sunscreen, the amount of product applied on volunteers, according to food and drug administration (FDA) and International protocols, is 2 mg/cm(2). However, different studies have shown that consumers actually apply much less product when exposed to the sun. Previous studies have reported contradictory findings in an attempt to correlate the amount applied in relation to SPF. The objective of the present study was to estimate the influence of the quantity of sunscreen applied in the determination of SPF, according to the FDA methodology. Subjects and methods Forty volunteers were included in two groups (SPF 15 and 30). The selected sunscreen was then applied in four different quantities (2, 1.5, 1.0 and 0.5 mg/cm(2)). All areas were irradiated with a solar simulator. After 24 h, the minimal erythemal dose (MED) and SPF were determined. Results In both groups, we observed that the SPF decreased when the amount of sunscreen applied was decreased. The differences between the 2 mg/cm(2) area and the others were significant in both groups (P < 0.001). The correlation between specified SPF and applied amount grew exponentially. Conclusion The protection provided by sunscreen is related to the amount of product applied. It is essential to educate consumers to apply larger amounts of sunscreen for adequate photoprotection.

Eating 'Green': Motivations behind organic food consumption in Australia

Central to the development of green lifestyles is the consumption of foods that by dint of their status as chemical-free, locally produced and/or free of genetically modified ingredients, reduce the environmental impact of food provision. Yet there are many other factors, such as health concerns, that may also encourage the consumption of 'green' foods. This paper explores the ways in which Australian consumers construct organic food-a sector of the food industry that is currently growing at between 20 and 50 percent per annum but is struggling to keep up with rising consumer demand. In order to examine the significance of 'green' signifiers in the consumption practices of Australian consumers a series of focus group interviews and a national consumer survey were conducted. These examined both those characteristics of food that were valued in general, and those meanings that were associated with organic food in particular. In very general terms, analysis reveals that while consumers believed organic foods to be healthy and environmentally sound-both of which were considered desirable-these characteristics were subsumed by an overarching concern with convenience. This does not mean that consumers did not hold genuinely positive environmental attitudes. Rather, it reflects a range of contradictory beliefs and practices that appeared to derive from the discursive conflict between conventional and organic food industries over environmental, health and safety claims. The paper concludes by identifying the barriers and opportunities for expanding the organic industry in Australia in the context of the ways organics is constructed by consumers.

The surplus value of Azorean macroalgae as health-promoting food. Protein, fiber, carbohydrates, lipids and vitamins determination

AICMA 2012 (BIT's 1st Annual International Congress of Marine Algae), World Expo Center, Dalian, China, 20-23 de Setembro.

Flavored waters: influence of ingredients on antioxidant capacity and terpenoid profile by HS-SPME/GC-MS

The antioxidant profiles of 39 water samples (29 flavored waters based on 10 natural waters) and 6 flavors used in their formulation (furnished by producers) were determined. Total phenol and flavonoid contents, reducing power, and DPPH radical scavenging activity were the optical techniques implemented and included in the referred profile. Flavor extracts were analyzed by HS-SPME/GC-MS to obtain the qualitative and quantitative profiles of the volatile fraction of essential oils. Results pointed out a higher reducing power (0.14 11.8 mg of gallic acid/L) and radical scavenging activity (0.29 211.5 mg Trolox/L) of flavored waters compared with the corresponding natural ones, an interesting fact concerning human health. Bioactive compounds, such as polyphenols, were present in all samples (0.5 359 mg of gallic acid/L), whereas flavonoids were not present either in flavored waters or in flavors. The major components of flavor extracts were monoterpenes, such as citral, R-limonene, carveol, and R-terpineol.

The influence of animation on functional food intake in children

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics

Determinació de diferents compostos bioactius en ingredients naturals per a l'elaboració de bombons funcionals amb beneficis cardiovasculars

Durant les darreres dècades, i degut, principalment, a un canvi en els hàbits alimentaris, hi ha hagut un augment a nivell mundial de malalties cròniques (l’obesitat, malalties cardiovasculars, etc.). En els països mediterranis hi ha menys incidència d’aquestes malalties i sembla ser que això es deu a l’anomenada dieta mediterrània. La dieta mediterrània es caracteritza per una combinació d’oli d’oliva com a grassa principal, verdures, hortalisses i fruites en abundància, lleguminoses, fruits secs, formatges i iogurt, peix, pa, pasta, cereals i els seus derivats i un consum moderat de vi i carns. Aquest model alimentari, ric en tocoferols, fitosterols i fitoestanols que ajuden a reduir el contingut de colesterol en sang, fa que en les poblacions mediterrànies hi hagi menys incidència de malalties cardiovasculars. Aquests compostos inhibeixen el deteriorament oxidatiu dels olis, actuen com agent antipolimerització per olis de fregir. Tenen capacitat de reduir els nivells de colesterol, evitant la incidència de malalties cardiovasculars. Els fitoesterols y fitoestanols es poden trobar en forma lliure o esterificada amb àcids grassos, àcids fenòlics i glucosa. Els objectius d’ aquest treball han estat, primer en el desenvolupament de mètodes d'anàlisi ràpids, fiables i robusts dels tocoferols, fitoesterols i fitoestanols i la seva aplicació en fruits sec, oli de segó, oli de pinyol de raïm i productes que els continguin. El primer mètode va estar basat en la cromatografía líquida (HPLC-DAD) amb extracció en fase sòlida (SPE) com tècnica alternativa a la saponificació para la determinació de fitoesterols lliures. Aquest mètode va estar aplicada a mostres de bombons que contenia fitoesterols. El segon mètode va estar basat en la cromatografia de gasos (GCFID) amb aponificació i SPE per quantificar fitoesterols i fitoestanols lliures, esterificats i totals. En els documents annexos es descriuen a profunditat els mètodes desenvolupats.

Sensory and Non-sensory Factors Behind the Liking and Choice of Healthy Food Products

A healthy and balanced diet can reduce health problems, such as overweight and metabolic syndrome. In general, people have a considerably good knowledge of what constitutes a healthy diet and how they could achieve it with their food choices. Besides, people argue that health is among their top five food choice motives. Nevertheless, the prevalence of overweight is increasing and other food choice motives, such as taste, seem to conflict with the health. Liking for food does not necessarily determine acceptance alone, thus several non-sensory factors, such as brand, country of origin and nutrition claim, can also influence. Moreover, consumers are individuals in how they prioritize sensory and nonsensory factors of foods, but e.g. increasing age, female gender and health concern have been connected to a more health-oriented dietary behaviour. To sum up, identifying different factors that can increase the liking and consumption of healthy food is essential in order to develop more attractive healthful food products. Adding vitamins, minerals, fibre or other ingredients to a food product can be used to enrich the nutritional quality of the products. However, this may be difficult in practice as regards the sensory quality and pleasantness of the foods. Generally, consumers are not willing to compromise on taste in food. On the other hand, consumers are very heterogeneous in their likings, and their personal values and attitudes may interact with preferences for specific sensory characteristics. The aims of this study were to investigate the effects of intrinsic product characteristics on sensory properties and hedonic responses; to determine the impact of few non-sensory factors; and to examine the interaction between sensory and non-sensory factors with consumers’ demographics, values and attitudes in liking of healthy model foods. The results showed that product composition influenced sensory quality and had an effect on hedonic responses. Adding flaxseed to bakery products showed a significant improvement in the nutritional quality without negative effects on sensory properties. On the other hand, the fortification of wellness beverages with vitamins and minerals may impart off-flavours. In general, sweetness of yoghurts, freshness of wellness beverages and low intensity of rye bread flavour appealed to consumers. Information about the domestic origin of yoghurts and claiming a specific function for wellness beverages enhanced liking. However, consumers who were more concerned about their health and considered natural content as an important food choice motive, rated sourer and less sweet yoghurts and wellness beverages as more pleasant. In addition, interest in health increased the consumption of rye breads and other whole grain breads among adolescents. The results showed that the optimal product quality in terms of intrinsic and extrinsic factors differs between individual consumers, and personal values and food choice motives can be connected to preferences for specific sensory characteristics of foods. This indicates that each food product needs to be considered in relation to its specific market niche, and to which segment of consumer will respond most positively to its characteristics.

Identifying and Characterizing New Ingredients in vitro for Prebiotic and Synbiotic use

The endogenous microbiota, constituting the microbes that live inside and on humans, is estimated to outnumber human cells by a factor of ten. This commensal microbial population has an important role in many physiological functions, with the densest microbiota population found in the colon. The colonic microbiota is a highly complex and diverse bacterial ecosystem, and a delicate balance exists between the gut microbiota and its host. An imbalance in the microbial ecosystem may lead to severe symptoms in and also beyond the gastrointestinal tract. Due to the important role of the gut microbiota in human health, means of its modification have been introduced in the dietary concepts of pro-, pre- and synbiotics. Prebiotics, which are usually carbohydrates, strive to selectively influence beneficial microbes resident in the colon with the aim of modifying the composition and functionality of the commensal microbial population towards a purportedly healthier one. The study of prebiotic effects on colonic micro-organisms is typically done by using human faecal material, though this provides relatively little information on bacterial populations and metabolic events in different parts of the colon. For this reason, several in vitro models have been developed to investigate the gut microbiota. The aim of this doctoral thesis was to screen through some of the promising prebiotic candidates, characterize their effects on the microbiota through the use of two in vitro methods (pure microbial cultures and a colon simulator model) and to evaluate their potential as emerging prebiotics or synbiotics when combined with the probiotic Bifidobacterium lactis . As a result of the screening work and subsequent colon simulation studies, several compounds with promising features were identified. Xylo-oligosaccharides (XOS), which have previously already shown promise as prebiotic compounds, were well fermented by several probiotic Bifidobacterium lactis strains in pure culture studies and in the following simulation studies utilizing the complex microbiota by endogenous B. lactis Another promising compound was panose, a trisaccharide belonging to isomalto-oligosaccharides (IMO) that also was also able to modify the microbiota in vitro by increasing the number of beneficial microbes investigated. Panose has not been widely studied previously and therefore, this thesis work provided the first data on panose fermentation in mixed colonic microbiota. Galacto-oligosaccharide (GOS) is an established prebiotic, and it was studied here in conjunction with another potential polygosaccharide polydextrose (PDX) and probiotic B. lactis Bi-07. In this final study, the synbiotics including GOS were more effective than the constituting pro- or prebiotics alone in modulating the microbiota composition, thus indicating a synergy resulting from the combination. The results obtained in this in vitro work can be, and have already been, utilized in product development aimed at the nutritional modification of the human colonic microbiota. Some of the compounds have entered the human clinical intervention phase to nvestigate in more detail the prebiotic and synbiotic properties seen in these in vitro studies.