Browsing by Author "Carvalho, Nelson Mota de"
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- Applied microbiology on poultry industry: from nutrition to the gut microbiotaPublication . Carvalho, Nelson Mota de; Oliveira, Diana Luazi; Costa, Célia Maria; Pintado, Manuela Estevez; Madureira, RaquelPoultry products (i.e., meat and eggs) are one of the major protein sources for the human diet. The animal’s diet is one of the key elements that the poultry industry has been focused on, to improve the animal’s performance, maintaining their healthy growth and, ultimately, high quality end products. The incorporation of functional ingredients in feed formulations, aiming to provide extra benefits and/or prevent diseases, has been considered efficient in maintaining the animal’s productivity and simultaneously ensure its well-being. Nutrient’s bioavailability varies throughout digestion and absorption within the poultry’s gastrointestinal tract (GIT). A reliable in vitro model, as the one developed and used in this study, capable of mimicking all digestion, absorptive and cecal fermentation processes, is a useful tool to study the potential benefits of feed supplemented with functional and/or bioactive ingredients. The developed in vitro gastrointestinal model simulates the chemical, enzymatic, and mechanical conditions prevailing in the chicken´s GIT, from beak to cecum. Fresh broiler’s cecal samples were used as inoculum for batch cecal fermentation and the impact of different feed formulations, on bacteria modulation, organic acids, and total ammonia nitrogen production, were assessed. Overall, this approach enables to evaluate, as close to reality as possible, the potential of target additives, providing a trustworthy tool for the development of functional feeds.
- Can supplemented skim milk (skm) boost your gut health?Publication . Carvalho, Nelson Mota de; Oliveira, Diana Luazi; Costa, Célia Maria; Pintado, Manuela; Madureira, Ana RaquelThe incorporation of functional ingredients, such as prebiotics and probiotics in food matrices, became a common practice in the human diet to improve the nutritional value of the food product itself. Worldwide, skim milk (SKM) is one of the most consumed food matrices, comprising all the essential nutrients desired for a balanced diet. Thus, the modulation of the human gut microbiota by SKM supplemented with different well-known functional ingredients was evaluated. Four well-studied prebiotics, fructo-oligosaccharides (FOS), galacto-oligosaccharides (GOS), mannanoligosaccharides (MOS) and inulin, and one probiotic product, UL-250® (Saccharomyces boulardii) were added at 1% (w/v) to SKM and subjected to a gastrointestinal in vitro model. The impact of each combination on gut microbiota profile and their fermentation metabolites (i.e., short-chain fatty acids–SCFA) was assessed by quantitative polymerase chain reaction (qPCR) and high-performance liquid chromatography (HPLC), respectively. The addition of FOS to SKM had promising results, showing prebiotic potential by promoting the growth of Lactobacillus, Bifidobacterium, and Clostridium cluster IV. Moreover, the increment of SCFA levels and the decrease of total ammonia nitrogen were observed throughout colonic fermentation. Overall, these results demonstrate that the combination SKM + FOS was the most beneficial to the host’s health by positively modulating the gut microbiota.
- A critical review on in vitro and ex vivo models of the intestinal epithelium of humans and monogastric animalsPublication . Costa, Célia Maria; Carvalho, Nelson Mota de; Oliveira, Diana Luazi de; Madureira, Ana RaquelRecently, the bioactive potential of several functional ingredients and biomolecules has been evaluated regarding human and animal nutrition. The digestive process from food intake to absorption and metabolism are important events that induce changes in ingredients, which affect their bioactivity. Consequently, there is a need to assess the bioavailability and bioaccessibility of these compounds. The methodology for the simulation of the human gastrointestinal tract has been standardized (INFOGEST protocol), while a gastrointestinal protocol for other animals (e.g., ruminants or broilers) has yet to be established. However, INFOGEST allows us only to predict bioaccessibility, leaving a gap regarding a methodology able to assess bioavailability by mimicking intestinal permeability and absorption. Several approaches—including in vitro, ex vivo, in situ and in vivo methods—can be found in the literature, aiming to tackle transepithelial routes, but leading to different results concerning the bioefficiency of the compounds studied. Therefore, this review aims to assess the current state-of-the-art regarding monogastric intestinal dynamics, absorption, and permeability events. Moreover, it compiled methodologies for simulating intestinal absorption in several biological systems, while reasoning their advantages, disadvantages, applications in ingredient development and the existing gaps.
- Development of a chicken gastrointestinal tract (GIT) simulation model: impact of cecal inoculum storage preservation conditionsPublication . Carvalho, Nelson Mota de; Costa, Célia Maria; Castro, Cláudia; Saleh, Mayra Anton Dib; Pintado, Manuela Estevez; Oliveira, Diana Luazi; Madureira, Ana RaquelA chicken gastrointestinal tract (GIT) simulation model was developed to help predict the potential effects of feed additives supplementation on chicken’ microbiota. The chemical and enzymatic conditions for oral, gastric, intestinal, and cecum fermentation phases were designed to closely resemble the chicken GIT conditions. For cecum fermentation, the inoculum was obtained from the cecal contents of 18 38-day broiler chickens. The impact of inoculum preservation on bacteria viability was assessed by comparing two methods of preservation with fresh inoculum: (1) 5% dimethyl sulfoxide (DMSO) at −80 °C and (2) 30% glycerol at −20 °C. The fermentation with fresh and frozen (DMSO method) inoculums was performed and compared using standard chicken feed (SCF) and SCF with 1% fructooligosaccharides (FOS), and inoculum control (IC) condition without feed matrix was used as a baseline. Inoculum’s viability was assessed throughout 90 days of storage by culture media platting, while bacterial growth and metabolites production during fermentation was evaluated by quantitative polymerase chain reaction (qPCR), high-performance liquid chromatography (HPLC), and total ammonia nitrogen quantification. The DMSO method was shown to be the most suitable for cecal inoculum storage. Higher growth of beneficial cecal bacteria for fresh inoculum was observed in SCF while for frozen inoculum, was the SCF + FOS condition. Also, frozen inoculum had lower activity of butyrate producers and proteolytic bacteria, showing different fermentation profiles. The GIT model developed showed to be useful to test the effect of feed additives supplementation.
- Do all prebiotics have the same impact on my gut microbiota? a case study using an in vitro gastrointestinal model to assess functional oligosaccharides added to milkPublication . Carvalho, Nelson Mota de; Costa, Célia Maria; Pintado, Manuela; Oliveira, Diana Luazi; Madureira, Ana Raquel
- Evaluation of mannans from saccharomyces cerevisiae as potential human microbiota modulatorsPublication . Faustino, Margarida; Carvalho, Nelson Mota de; Durão, Joana; Pintado, Manuela E.; Madureira, Ana Raquel; Carvalho, Ana P.
- Fermented foods and beverages inhuman diet and their influence on gut microbiota and healthPublication . Carvalho, Nelson Mota de; Costa, Eduardo M.; Silva, Sara; Pimentel, Lígia; Fernandes, Tito H.; Pintado, Manuela EstevezDietary changes have accompanied the evolution of humanity and is proven to be fundamental in human evolution and well-being. Nutrition is essential for survival and as a matter of health and equilibrium of the human body. About 1/3 of the human diet is composed by fermented foods and beverages, which are widely distributed and consumed in different societies around the world, no matter the culture and lifestyle. Fermented foods are derived from the fermentation process of different substrates by microorganisms, and more importantly to humans, by those with beneficial characteristics, due to the positive impact on health. Food is transformed in the gut, gaining new proprieties, and increasing its value to the organism. The effects of fermented foods and beverages can be assessed by its influence at the gut microbiota level. Recent studies show the major importance of the gut microbiota role in modulating the organism homeostasis and homeorhesis. More crosslinks between health, gut microbiota and diet are being established especially in the gut–brain axis field. Therefore, the benefits of diet, in particularly of fermented foods and beverages, should be studied and pursued in order to promote a good health status.
- Human and animal in vitro gastrointestinal models: importance and applicationsPublication . Carvalho, Nelson Mota de; Oliveira, Diana Luazi; Costa, Célia Maria; Pintado, Manuela Estevez; Madureira, Ana Raquel
- Impact of novel clean label ham formulations on the human gut microbiotaPublication . Carvalho, Teresa Bento de; Barbosa, Joana Bastos; Carvalho, Nelson Mota de; Souza, Carla; Costa, Célia; Komora, Norton; Azevedo, Alexandra; Madureira, Ana Raquel; Teixeira, PaulaIntroduction: The influence of diet on the human gut microbiota has been the subject of much debate, particularly in relation to innovative products that are still considered novel. Dietary habits have a fundamental impact on the human gut microbiota, which explains the variations observed between individuals and over the course of a lifetime. The aim of this study was to evaluate the effect of four clean label ham formulations (Table 1) (without sodium nitrite and with natural nitrate sources combined with the addition of nitrate reducing cultures) on the human gut microbiota of potential consumers after in vitro digestion, according to the INFOGEST protocol, and colonic fermentation. Methods: DNA extraction from the stored pellets was carried out using the Invitrogen PureLink™ Microbiome DNA Purification Kit. 16S rRNA gene sequence data was processed with QIIME2 by Novogene UK; Quantification of fermentation metabolites, short-chained fatty acids and branched- chained fatty acids, by high-performance liquid chromatography (HPLC) from supernatant. The impact of each novel formulation and for a faecal inoculum control (Inoc) on the gut microbiota profile and fermentation metabolites (i.e., SCFAs) was assessed by next-generation sequencing (NGS) and high-performance liquid chromatography (HPLC), respectively. Results and discussion: Higher concentration of SCFA (Acetate + Butyrate + Propionate) after 48 h colonic fermentation were found for: D (65.96 mM) > A (63.36 mM) > E (59.35 mM) > C (58.64 mM) > B (57.22 mM) > Inoc (16.20 mM); Lowest SCFA concentrations were found for the inoculum control (Inoc), as expected, due to the lack of nutrients to promote bacterial fermentation over time. The most abundant phylum present were Bacillota and Pseudomonadota for all samples (A, B, C, D, E and Inoc) and all time points (0h, 24h and 48h); Higher relative abundance of Pseudomonadota was observed for all samples when compared to the inoculum control (Inoc); After fermentation, relative abundance of Bacteroidota is higher for the ham control sample (added sodium nitrite) than for the samples that are formulated with natural plant nitrate coupled with starter cultures. Conclusions: There were no discernible variations in SCFA levels or microbial populations during colonic fermentation between the new formulations and the conventional ham, indicating that the suggested clean label approach produced encouraging outcomes; Additional investigation should provide light on the endogenous production of volatile and non-volatile chemicals (nitrosamines) in vitro, their effects on the microbiota in the human gut, and any potential toxicity to human intestinal epithelial cells.
- Importance of gastrointestinal in vitro models for the poultry industry and feed formulationsPublication . Carvalho, Nelson Mota de; Oliveira, Diana Luazi; Saleh, Mayra Anton Dib; Pintado, Manuela Estevez; Madureira, Ana RaquelThe animal’s diet is a crucial factor, as poultry feed formulations influences greatly their development, well-being and final products quality, i.e., meat and eggs. Therefore, the search for feed additives that provide concomitantly better performances, low-cost usage, guarantying the animal well-being and products safety, became a priority to the poultry industry. Although in an early stage, research has been focused on developing the optimal cost-efficient feed formulations, taking into account the chicken’s physiology and function of the gastrointestinal tract and intestinal microbiota. This review discusses a number of concepts and novel approaches towards the optimization of poultry’s feed formulations, by critically encompassing the animal’s growth and performance. Additionally, it highlights the in vitro gastrointestinal models capabilities as a potential solution to test highly nutritive, well-balanced and efficient feed formulations within a circular economy framework.
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