Minimum inhibitory concentrations (MIC) assays are often questioned for their representativeness. Especially when foodborne pathogens are tested, it is of crucial importance to also consider parameters of the human digestive system. Hence, the current study aimed to assess the inhibitory capacity of two antibiotics, ciprofloxacin and tetracycline, against Salmonella enterica and Listeria monocytogenes, under representative environmental conditions. More specifically, aspects of the harsh environment of the human gastrointestinal tract (GIT) were gradually added to the experimental conditions starting from simple aerobic lab conditions into an in vitro simulation of the GIT. In this way, the effects of parameters including the anoxic environment, physicochemical conditions of the GIT (low gastric pH, digestive enzymes, bile acids) and the gut microbiota were evaluated. The latter was simulated by including a representative consortium of selected gut bacteria species. In this study, the MIC of the two antibiotics against the relevant foodborne pathogens were established, under the previously mentioned environmental conditions. The results of S. enterica highlighted the importance of the anaerobic environment when conducting such studies, since the pathogen thrived under such conditions. Inclusion of physicochemical barriers led to exactly opposite results for S. enterica and L. monocytogenes since the former became more susceptible to ciprofloxacin while the latter showed lower susceptibility towards tetracycline. Finally, the inclusion of gut bacteria had a bactericidal effect against L. monocytogenes even in the absence of antibiotics, while gut bacteria protected S. enterica from the effect of ciprofloxacin.

Protein derived from chia (Salvia hispanica L.), characterized by a balanced amino acid composition, represents a potentially healthier and environmentally friendly alternative poised for innovation within the plant-based food sector. It was hypothesized that the growing location of chia seeds and processing techniques used might influence protein digestion patterns, which in turn could affect the biological functions of the digestion products. To examine this hypothesis, we assessed the gastrointestinal fate of degummed-defatted flour (DDF), protein concentrate (PC), and isolated albumin (Alb) and globulin (Glo) fractions. Furthermore, we compared the antioxidant and anti-inflammatory activities of the resulting digesta by means of in vitro and cellular assays. Post-gastrointestinal digestion, the PC exhibited elevated levels of soluble protein (7.6 and 6.3 % for Mexican and British PC, respectively) and peptides (24.8 and 27.9 %, respectively) of larger molecular sizes compared to DDF, Alb, and Glo. This can be attributed to differences in the extraction/fractionation processes. Leucine was found to be the most prevalent amino acids in all chia digesta. Such variations in the digestive outcomes of chia protein components significantly influenced the bioactivity of the intestinal digestates. During gastrointestinal transit, British Glo exhibited the best reactive oxygen species (ROS) inhibition activity in oxidative-stressed RAW264.7 macrophages, while Mexican digesta outperformed British samples in terms of ROS inhibition within the oxidative-stressed Caco-2 cells. Additionally, both Mexican and British Alb showed effectively anti-inflammatory potential, with keratinocyte chemoattractant (KC) inhibition rate of 82 and 91 %, respectively. Additionally, Mexican PC and Alb generally demonstrated an enhanced capacity to mitigate oxidative stress and inflammatory conditions in vitro. These findings highlight the substantial potential of chia seeds as functional food ingredients, resonating with the shifting preferences of health-conscious consumers.

Natural waxes have demonstrated exceptional potential as oil gels for saturated and trans fatty acids, but their application has been limited by issues such as temperature sensitivity, lack of stability and durability, and compatibility. In this study, three types of wax (Beeswax (BW), Rice bran wax (RBW), and Carnauba wax (CW)) were combined with calcium hydroxide to produce calcified wax. The calcified Korean pine seed oil gel obtained by heating and stirring with Korean pine seed oil is responsive to temperature and has environmental adaptability. The effects of critical gel concentration, temperature regulation, texture properties, microstructure, oil-holding capacity, and FT-IR on the quality parameters of oil gel were investigated. Additionally, an in vitro digestion model was developed to comprehend the decomposition rate of fat during gel structure digestion and transportation. The results demonstrated a close correlation between the critical gelation concentration and calcium ion content. Furthermore, after calcification, the hardness followed the order BW > CW > RBW. Moreover, there was an approximate 10 °C increase in wax melting point. Conversely, BW:Ca exhibited the lowest oil leakage. The microstructures revealed that the oil gels formed post-wax calcification exhibited similar fractal dimension (Db) values (<7 μm), and the intermolecular forces were characterized by van der Waals forces, which were consistent with those observed in the non-calcified group. In conjunction with the vitro digestion simulation, our findings demonstrated that RBW and CW oil gels gradually released 20%, 35%, and 35% of free fatty acids (FFA) within the initial 30 min of intestinal digestion. Importantly, the FFA release rate was significantly attenuated, thereby providing a foundation for developing wax-based gel processed foods that facilitate gentle energy release benefits for healthy weight management.

Essential oil (EO) nanoemulsions have been recently studied due to their antimicrobial properties. Nevertheless, little is known about their possible negative effect against human gut microorganisms during their passage though the gastrointestinal tract. This work studied the effect of digestible (corn oil) or non-digestible (paraffin oil) citral nanoemulsions against specific microorganisms of human microflora under in vitro digestion conditions. The use of a citral lipid carrier (paraffin oil or corn oil) decreased the nanoemulsion particle size and increased its stability after gastric conditions with regards to the pure citral nanoemulsions. Digestible nanoemulsions formulated with corn oil and citral presented a lower bactericidal activity against Lactobacillus acidophilus and Escherichia coli after being subjected to in vitro digestion conditions in comparison to the initial nanoemulsion. However, a non-digestible nanoemulsion formulated with paraffin oil and citral presented a similar antimicrobial activity against L. acidophilus and E. coli to the one of the initial nanoemulsion. This evidences that non-digestible nanoemulsions may entrap the citral in the lipid core and thus retaining its antimicrobial potential during their passage though the gastrointestinal tract. Hence, this work evidences the impact of the lipid carrier digestibility when formulating antimicrobial nanoemulsions on certain intestinal probiotic bacteria.

Transglutaminase (TGase) is gaining increasing recognition as a novel and healthier bio-binder for meat analogs. This work focused on the TGase-induced crosslinking behaviors, and then evaluated the difference in quality characteristics (Texture, water distribution, cooking properties, volatile flavor and protein digestibility) of peanut protein-based burger patties treated with TGase and traditional binder (methylcellulose, MC). TGase-catalyzed crosslinking, enabling amino acids to participate in the formation of covalent bonds rather than non-covalent bonds, and promoted the formation of protein aggregates and dense gel networks by changing the protein structure, ultimately improving the quality characteristics of burger patties. Compared with the TGase treatment, MC-treated burger patties showed a greater texture parameter, lower cooking loss, higher flavor retention but a lower degree of digestibility. The findings will contribute to a better understanding of the roles of TGase and traditional binders in plant-based meat analogs.

This thorough study analyses the amounts of 43 minerals and trace elements in non-traditional wheat grains, flakes, and undigested flake portions using ICP-MS and establishes declines in their respective contents after the flake production. It also identifies appropriate dietary intakes, in vitro digestibility values, retention factors, and metal pollution indexes. The element contents in wheat flakes are lower than in wheat grains after the hydrothermal treatment process, and their declines are: Na (48-72%), Ce (47-72%), Sr (43-55%), Tl (33-43%), Ti (32-41%), U (31-44%), Ho (29-69%), Cr (26-64%), Zr (26-58%), Ag (25-52%), and Ca (25-46%). The flakes significantly contributed to the recommended dietary intake or adequate intake of particular elements for men of all categories as follows: Mn (143%) > Mo > Cu > Mg ≥ Cr > Fe (16%); for women: Mn (up to 183%) > Mo > Cu > Cr ≥ Mg > Fe (7-16%); for pregnant women aged 19-30: Mn (165%) > Mo > Cu > Mg > Cr (25%); and finally, for lactating women: Mn (127%) > Mo > Cu > Mg > Cr (17%). The contributions to the provisional tolerable weekly or monthly intakes of all toxic elements were established as being within the official limits. The daily intakes for non-essential elements were also calculated. The retention factors were calculated to assess the element concentrations in the undigested part using the digestibility values (87.4-90.5%). The highest retention factors were obtained for V (63-92%), Y (57-96%), Ce (43-76%), Pb (34-58%), Tl (32-70%), Ta (31-66%), and Ge (30-49%). K, Mg, P, Zn, Ba, Bi, Ga, Sb, Cu, Ni, and As appear to be released easily from flake matrices during digestion. The metal pollution index has been confirmed as being lower for non-traditional wheat flakes when compared with grains. Importantly, 15-25% of the metal pollution index assessed for native flakes remains in the undigested flake portion after in vitro digestion.

This experiment evaluated the difference between computer-controlled simulated digestion and in vivo stomach-small intestinal or large intestinal digestion for growing pigs. Five diets including a corn-soybean meal basal diet and four experimental diets with rapeseed meal (RSM), cottonseed meal (CSM), sunflower meal (SFM), or peanut meal (PNM) were assigned to each group of five barrows installed terminal ileal cannula or distal cecal cannula in a 5 × 5 Latin square design. Ileal digesta and feces were collected for the determination of digestibility of dry matter (DM) and gross energy (GE) as well as digestible energy (DE) at terminal ileum and total tract. The large intestinal digestibility and DE were calculated by the difference between measurements obtained at the terminal ileum and those obtained from total tract. In vitro stomach-small intestinal digestibility and DE for diets and plant protein meals were determined by stomach-small intestinal digestion in a computer-controlled simulated digestion system (CCSDS). The in vitro large intestinal digestibility and DE of diets were determined in a CCSDS using ileal digesta and enzymes extracted from cecal digesta of pigs. The in vitro large intestinal digestibility and DE of four plant protein meals were determined by the difference between stomach-small intestinal and total tract digestion in the CCSDS. For the experimental diets, the in vitro ileal digestibility and DE were not different from corresponding in vivo values in basal diet and PNM diet, but greater than corresponding in vivo values for diets with RSM, CSM, and SFM (P < 0.05). No difference was observed between in vitro and in vivo large intestinal digestibility and DE in five diets. For the feed ingredients, the in vitro ileal digestibility and DE did not differ from corresponding in vivo ileal values in RSM and PNM but were greater than the in vivo ileal values in CSM and SFM (P < 0.05). The in vitro large intestinal GE digestibility and DE were not different from in vivo large intestinal values in RSM, CSM, and PNM, but lower than in vivo large intestinal values in SFM. This finding may relate to the higher fiber content of plant protein meals resulting in shorter digestion time of in vivo stomach-small intestine thus lower digestibility compared to in vitro, indicating it is necessary to optimize in vitro stomach-small intestinal digestion time.
Comparable in vitro and in vivo values are crucial to develop a novel in vitro digestion technique for growing pigs. The current study evaluated the difference between computer-controlled simulated digestion and in vivo stomach–small intestinal or large intestinal digestion for growing pigs. Five diets including a corn–soybean meal basal diet and four experimental diets with rapeseed meal (RSM), cottonseed meal (CSM), sunflower meal (SFM), or peanut meal (PNM) were used to compare the in vitro and in vivo digestion. Our study demonstrated that the in vitro ileal digestibility of energy was not different from corresponding in vivo values in basal diet and PNM diet, but greater than corresponding in vivo values for diets with RSM, CSM, and SFM. The in vitro stomach–small intestinal digestibility was greater than in vivo digestibility, resulting in less digestible substrates hydrolyzed by in vitro large intestinal fluid, whereas more digestible substrates can be digested by in vivo large intestine in plant protein meals. This difference may relate to the higher fiber content of plant protein meals resulting in shorter digestion time of in vivo stomach–small intestine thus lower digestibility compared to in vitro. Therefore, it is necessary to optimize in vitro stomach–small intestinal digestion time.

Cellular pulse ingredients are increasingly being studied but little knowledge on their proteolysis patterns upon digestion is available. This study investigated a size exclusion chromatography (SEC) approach to study in vitro protein digestion in chickpea and lentil powders, providing novel insights into proteolysis kinetics and the evolution of molecular weight distributions in the (solubilized) supernatant and (non-solubilized) pellet fractions. For the quantification of proteolysis, SEC-based analysis was compared to the commonly used OPA (o-phthaldialdehyde) approach and nitrogen solubilized upon digestion, leading to highly correlated proteolysis kinetics. Generally, all approaches confirmed that microstructure dictated proteolysis kinetics. However, SEC analysis delivered an additional level of molecular insight. For the first time, SEC revealed that while bioaccessible fractions reached a plateau in the small intestinal phase (around 45-60 min), proteolysis continued in the pellet, forming smaller but mostly insoluble peptides. SEC elutograms showed pulse-specific proteolysis patterns, unidentified using other current state-of-the-art methods.

In this study an in vitro static digestion method mimicking the elderly gastrointestinal conditions was designed by adapting the physiological parameters described in the INFOGEST standardized static in vitro digestion protocol, i.e., pH, digestive phase duration, concentrations of enzymes and bile salts, to the aged GI transit. The digestibility of proteins from different sources (pea, rice, wheat, and milk whey) was then assessed. Protein digestive behaviour was monitored after gastric and intestinal phases by BCA assay and SDS-PAGE to assess protein hydrolysis both from a quantitative and a qualitative point of view. Digested samples were also analysed for physical characteristics in terms of particle size and zeta potential. Data acquired under elderly gastrointestinal conditions were compared to those obtained by using the INFOGEST protocol designed to study adult digestion. Results clearly showed that the elderly gastrointestinal conditions deeply affected proteolysis leading to a general reduction of protein digestibility in comparison to the adult model. The proteolysis extent depended on the protein source with whey and rice proteins showing about 20% reduction using the model mimicking the elderly gut, followed by pea (about 10% reduction) and wheat (about 4% reduction) proteins. The knowledge of protein digestibility under elderly gastrointestinal conditions generated in this study could be useful in the attempt to develop age-tailored products.

Curcumin-loaded lipid-based nano delivery systems (nanoemulsions-NE, solid lipid nanoparticles-SLN and nanostructured lipid carriers-NLC) were subjected to different food simulants to evaluate curcumin\'s in vitro release kinetics and particlés stability. The nano delivery systems were also incorporated into a model beverage and their physicochemical properties were evaluated during storing period. Curcumin\'s bioaccessibility of beverages containing nano delivery systems were assessed through an in vitro digestion process. All nano delivery systems showed a higher curcumin\'s release and lower particle stability at 50 % ethanol (lipophilic food simulant) comparatively to hydrophilic food simulants. NLC and SLN showed a good particle\'s stability within the beverage during storing period, while NE presented high instability immediately after incorporation in the beverage. NLC and SLN did not affect beverage\'s stability relatively to pH, however the beverage with NLC was slightly more stable regarding color. Beverage with SLN presented higher curcumin bioaccessibility compared to the beverage with NLC, however it showed lower curcumin\'s stability.