Functional components and lipid quality assessment in vegetable foods: analytical and technological approaches

Lipids are important components that contribute very significantly to nutritional and technological quality of foods because they are the least stable macro-components in foods, due to high susceptibility to oxidation. When rancidity take place, it makes food unhealthy and unacceptable for consumers. Thus, the presence of antioxidants, naturally present of added to foods, is required to enhance shelf life of foods. Moreover, antioxidant like phenolic compounds play an important role in human health enhancing the functionality of foods. The aim of this PhD project was the study of lipid quality and lipid oxidation in different vegetable foods focusing on analytical and technological aspects in order to figure out the effects of lipid composition and bioactive compounds (phenolic compounds, omega-3 fatty acids and dietary fiber) addition on their shelf life. In addition, bioavailability and antioxidant effects of phenolic compounds in human and animals, respectively, were evaluated after consumption of vegetable foods. The first section of the work was focused on the evaluation of lipid quality impact on technological behaviour of vegetable foods. Because of that, cocoa butter with different melting point were evaluated by chromatographic techniques (GC, TLC) and the sample with the higher melting point showed the presence of fatty acids, triglycerides, 2-monoglycerides and FT-IR profile different from genuine cocoa butter, meaning an adding of foreign fat (lauric-fat) not allowed by the law. Looking at lipid quality of other vegetable foods, an accelerated shelf life test (OXITEST®), was used to evaluate of lipid stability to oxidation in tarallini snacks made up using different lipid matrices (sunflower oil, extravirgin olive oil and a blend of extravirgin olive oil and lard). The results showed a good ability of OXITEST® to discriminate between lipid unsaturation and different cooking times, without any samples fat extraction. In the second section, the role of bioactive compounds on cereal based food shelf life was studied in different bakeries by GC, spectrophotometric methods and capillary electrophoresis. It was examined the relationships between phenolic compounds, added with flour, and lipid oxidation of tarallini and frollini. Both products showed an increase in lipid oxidation during storage and antioxidant effects on lipid oxidation were not as expected. Furthermore, the influence of enrichment in polyunsaturated fatty acids on lipid oxidation of pasta was evaluated. The results proved that LC n-3 PUFA were not significantly implicated in the onset of oxidation in spaghetti stored under daylight and accelerated oxidation in a laboratory heater. The importance of phenolic compounds as antioxidant in humans and rats was also studied, by HPLC/MS in the latter section. For this purpose, apigenin and apigenin glycosides excretion was investigated in six women’s urine in a 24 hours study. After a single dose of steamed artichokes, both aglicone and glucuronide metabolites were recovered in 24 h urine. Moreover, the effect of whole grain durum wheat bread and whole grain Kamut® khorasan bread in rats were evaluated. Both cereals were good sources of antioxidants but Kamut® bread fed animals had a better response to stress than wheat durum fed, especially when a sourdough bread was supplied.

Wheat and low-input agriculture: agronomic, nutritional and nutraceutical implications

Recently, the increasing interest in organic food products and environmental friendly practices has emphasized the importance of selecting crop varieties suitable for the low-input systems. Additionally, in recent years the relationship between diet and human health has gained much attention among consumers, favoring the investigations on food nutraceutical properties. Among cereals, wheat plays an important role in human nutrition around the world and contributes to the daily intake of essential nutrients such as starch and protein. Moreover, whole grain contains several bioactive compounds that confer to wheat-derived products unique nutraceutical properties (dietary fibre, antioxidants). The present research provided interesting insights for the selection of wheat genotypes suitable for low-input systems and the development of specific breeding programs dedicated to organic farming. The investigation involved 5 old not dwarf genotypes (Andriolo, Frassineto, Gentil rosso, Inallettabile, Verna) and 1 modern dwarf variety (Palesio), grown under biodynamic management, over two consecutive growing seasons (2009/2010, 2010/2011). Results evidenced that under low-input farming some investigated old wheat genotypes (Frassineto, Inallettabile) were comparable to the modern cultivar in terms of whole agronomic performance. As regards the nutritional and nutraceutical properties, some old genotypes (Andriolo, Gentil rosso, Verna) emerged for their relevant content of several investigated phytochemicals (such as insoluble dietary fibre, polyphenols, flavonoids, in vitro antioxidant activity) and nutrients (protein, lipid, minerals). Despite of the low technological features, the six wheat varieties grown under low-input management may efficiently provide raw material for the preparation of traditionally processed bread with valuable sensory and nutritional properties. Results highlighted that old wheat varieties have peculiar phytochemical composition and may be a valuable source of nutraceutical compounds. Some of the genetic material involved in the present study may be used in breeding programs aimed at selecting varieties suitable for low-input farming and rich in health-promoting compounds.

Possibilities for the healthy and nutritional improvement of confectionery and sweet products.

The overall objective of this PhD was to investigate the possibility to increase the nutritional value of confectionary products by the use of natural ingredients with healthy functions. The first part of the thesis focused on the possible substitution of the most characteristic component of confectionary products, i.e. refined sugar. Many natural whole sweetening alternatives are available, though not widely used; the use of molasses, the byproduct of sugar beet and cane production, still rich in healthy components as minerals and phytochemicals is hereby discussed; after having verified molasses effectiveness in oxidative stress counteraction on liver cultured cells, the higher antioxidant capacity of a sweet food prepared with molasses instead of refined sugar was confirmed. A second step of the project dealt with another main ingredient of various sweet products, namely wheat. Particularly, the exploitation of soft and durum wheat byproducts could be another sustainable strategy to improve the healthy value of confectionery. The isolation of oligosaccharides with bioactive functions form different fractions of the wheat milling stream was studied and the new ingredients were shown to have a high dietary fiber and antioxidants content. As valid alternative, product developers should consider the appealing and healthy addition of ancient grains flour to sweet baked goods. The possibility of substituting the modern whole durum wheat with the ancient Kamut® khorasan was considered, and the antioxidant and anti-inflammatory effects of these grains were evaluated and compared both in vitro and in vivo on rats. Finally, since high consumption of confectionery is a risk factor for obesity, a possible strategy for the counteraction of this disease was investigated. The ability of three bioactives in inhibiting adipocytes differentiation was investigated. In fact, theoretically, compounds able to influence adipogenesis could be used in the formulation of functional sweet products and contribute to prevent obesity.

Gut microbiota perturbation induced by emerging dietary habits: an in vitro colon model to predict their effects by a microbiomic and metabolomic approach

Over the past 30 years, unhealthy diets and lifestyles have increased the incidence of noncommunicable diseases and are culprits of diffusion on world’s population of syndromes as obesity or other metabolic disorders, reaching pandemic proportions. In order to comply with such scenario, the food industry has tackled these challenges with different approaches, as the reformulation of foods, fortification of foods, substitution of ingredients and supplements with healthier ingredients, reduced animal protein, reduced fats and improved fibres applications. Although the technological quality of these emerging food products is known, the impact they have on the gut microbiota of consumers remains unclear. In the present PhD thesis, the recipient work was conducted to study different foods with the substitution of the industrial and market components to that of novel green oriented and sustainable ingredients. So far, this thesis included eight representative case studies of the most common substitutions/additions/fortifications in dairy, meat, and vegetable products. The products studied were: (i) a set of breads fortified with polyphenol-rich olive fiber, to replace synthetic antioxidant and preservatives, (ii) a set of Gluten-free breads fortified with algae powder, to fortify the protein content of standard GF products, (iii) different formulations of salami where nitrates were replaced by ascorbic acid and vegetal extract antioxidants and nitrate-reducers starter cultures, (iv) chocolate fiber plus D-Limonene food supplement, as a novel prebiotic formula, (v) hemp seed bran and its alkalase hydrolysate, to introduce as a supplement, (vi) milk with and without lactose, to evaluate the different impact on human colonic microbiota of healthy or lactose-intolerants, (vii) lactose-free whey fermented and/or with probiotics added, to be introduced as an alternative beverage, exploring its impact on human colonic microbiota from healthy or lactose-intolerants, and (viii) antibiotics, to assess whether maternal amoxicillin affects the colon microbiota of piglets.

Different dietary fibres included in the gestation and lactation diet shape sow's microbiota, modify colostrum and milk composition and affect piglet growth and health

This thesis reports three experimental studies that may contribute to understand how the sources or types of dietary fibres (DFs) included in sow diet with similar level of total DFs influence the composition of colostrum and milk and their related effects on offspring performance and gut microbiota. The first study showed that decreasing the level of hemicelluloses (HCs) in sow’s lactation diet increased the proportion of butyrate and the concentration of volatile fatty acids (VFAs), copper and threonine in milk. Simultaneously, the post-weaning growth of low birthweight piglets was improved, and the diarrhoea occurrence was reduced during the second week post-weaning. The second study showed that the level of HCs in the diet of lactating sows affected their faecal microbiota, modified the VFA profile in sow’s faeces during lactation and barely impacted the faecal microbiota of slow and fast growing piglets. The third study showed that replacing a source soluble DFs by one of insoluble DFs in sow’s diet during late gestation and lactation reduced farrowing duration, increased total VFAs and lactoferrin concentrations in colostrum, improved growth performance from birth to 1 day of lactation, during the post-weaning period and throughout the study, and reduced diarrhoea occurrence during the first week post-weaning. Finally, a fourth study proposed a workflow to analyse low biomass samples from the umbilical cord blood aiming at investigating the existence of a pre-birth microbiota with no substantial findings to confirm this hypothesis. Overall, the results of these studies confirmed that, besides the level of DFs, the sources, and the types of DFs included in the sow's diet shape the sow's microbiota, influence the composition of colostrum and milk, and improve offspring performance, but with limited impacts on the microbiota of piglets.