Two experiments were carried out to study the effects of dietary supplementation with the garlic ... more Two experiments were carried out to study the effects of dietary supplementation with the garlic (Allium sativum)-derived product propyl propane thiosulfonate (PTS-O) on the intestinal log 10 number of copies of enteropathogens in broiler chickens, together with their intestinal morphology and growth performance. The additive had no significant effect on feed intake at any dose assayed. In experiment 1 (1 to 21 d of age), the BW of chickens fed on 45 mg of PTS-O/ kg of diet was higher (P < 0.01) than that of controls. Birds fed on diets containing 45 and 90 mg of PTS-O/ kg of diet had improved (P < 0.01) feed:gain ratios compared with the controls at 21 d of age. Ileal villus height, width and surface area, mucosal thickness, and muscular layer thickness were considerably greater (P < 0.01) than control values in chickens fed 90 mg of PTS-O/kg of diet. The Clostridium perfringens log 10 number of counts was not significantly affected at any dose assayed.
Castrated male Iberian (n = 12) and Landrace × Large White (n = 12) pigs were used to study histo... more Castrated male Iberian (n = 12) and Landrace × Large White (n = 12) pigs were used to study histological structure and enzymatic activity in the small intestine at 3 points of the productive cycle (BW = 15, 50, and 115 kg). Both strains were fed the same cereal-based diets (DE = 2,799 kcal•kg −1 , and CP = 15%) throughout the entire experimental period. Differences (P < 0.05) in histometrical variables (villus height, width and surface, crypt depth, villus height/ crypt depth relationship, mucosal thickness, muscular layer thickness, and number of goblet cells) were found among samples of small intestinal sections (duodenum, jejunum, and ileum) at the 3 productive stages studied. Also, differences (P < 0.05) in histometrical variables of small intestinal samples were found between Iberian and lean pigs at all productive stages, although these differences tended to disappear with age. Differences (P < 0.05) in enzymatic activities (lactase, sucrase, maltase, isomaltase, aminopeptidase, and alkaline phosphatase) of small intestinal samples were found between the different intestinal sections at all productive stages studied. Although differences (P < 0.05) in enzymatic activities of small intestinal samples were found between Iberian and lean pigs at all productive stages, values tended to equalize with pig age. We concluded that differences previously found in dietary nutritional utilization between Iberian and lean strains are likely not due to differences in intestinal absorption or hydrolytic capacity.
Cowpea (Vigna unguiculata) value as a functional food is still quite underexplored. Weaned Wistar... more Cowpea (Vigna unguiculata) value as a functional food is still quite underexplored. Weaned Wistar albino rats were fed one of four experimental equilibrated diets based on casein or cowpea meal as the only protein sources. RT-qPCR analysis showed that the replacement of casein for raw or treated cowpea meal lowered Escherichia/Shigella faecal counts and increased Lactobacilli, Bifidobacteria, Bacteroides, Clostridium leptum/Ruminococcus, and Faecalibacerium prausnitzii log 10 counts. Sequencing (Illumina) analysis confirmed these results and revealed that rats fed the cowpea-based diets presented higher proportions of potentially beneficial bacterial groups. Non-starch polysaccharides digestibility was associated with bacterial groups related with the control diet, while oligosaccharides digestibility was associated with cowpea diets. In conclusion, the inclusion of Vigna unguiculata in diets for rats, particularly in the untreated form, induced a healthier intestinal microbiota composition, which was mainly linked to the fermentation of oligosaccharides of the raffinose family.
Journal of the Science of Food and Agriculture, Jul 8, 2013
BACKGROUND: Legume seed proteins have to be chemically characterized in order to properly link th... more BACKGROUND: Legume seed proteins have to be chemically characterized in order to properly link their nutritional effects with their chemical structure. RESULTS: Vicilin and albumin fractions devoid of cross-contamination, as assessed by mass peptide fingerprinting analysis, were obtained from defatted pea (Pisum sativum cv. Bilbo) meal. The extracted protein fractions contained 56.7-67.7 g non-starch polysaccharides kg −1. The vicilin fraction was higher than legumins in arginine, isoleucine, leucine, phenylalanine and lysine. The most abundant amino acids in the albumin fraction were aspartic acid, glutamic acid, lysine and arginine, and the amounts of methionine were more than double than those in legumins and vicilins. The pea albumin fraction showed a clear enrichment of protease inhibitory activity when compared with the seed meal. In vitro digestibility values for pea proteins were 0.63 ± 0.04, 0.88 ± 0.04 and 0.41 ± 0.23 for legumins, vicilins and albumins respectively. CONCLUSION: Vicilin and albumin fractions devoid of cross-contamination with other proteins were obtained from pea seed meal. The vicilin fraction also contained low amounts of soluble non-starch polysaccharides and was enriched in isoleucine, leucine, phenylalanine and lysine. In vitro digestibility values for pea proteins were similar or even numerically higher than those for control proteins.
The effects of dietary sweet lupin (Lupinus angustifolius, Unicrop) seed meal or its insoluble fi... more The effects of dietary sweet lupin (Lupinus angustifolius, Unicrop) seed meal or its insoluble fiber (nonstarch polysaccharides + lignin) on performance, digestibility and nitrogen utilization in growing rats were studied in four experiments. Globulin proteins isolated from lupin, faba bean (Vicia faba L. minor) or soybean (Glycine max) were also incorporated into purified diets as replacements for lactalbumin (control) and the nutritional effects were evaluated. Isocaloric, legume-based diets supplemented with amino acids were used. Final weight gain, gain:feed ratios, nitrogen retention and net protein utilization of the animals fed whole lupin meal-based diets for 10 d were inferior to those of controls. In contrast, adding lupin insoluble fiber to a control diet produced no adverse effects. Ileal starch and apparent nitrogen digestibilities, and fecal digestibility of starch in lupin-fed rats were higher than those of controls, but fecal true nitrogen digestibility was lower. Replacement of lactalbumin with globulin proteins from lupin or faba bean depressed food intake and protein utilization, but only performance was affected by consumption of soybean globulins. Rats consuming lupin or faba bean globulins excreted significantly more nitrogen, particularly as urea through the urine. This did not occur in rats fed soybean globulins. Urea concentration in plasma was higher in rats fed diets containing lupin meal or legume globulins. The concentrations of urea, arginine and ornithine in plasma increased significantly compared with control values after 3 to 9 h of a lupin diet. After 9 h, plasma lysine was also decreased. We concluded that the main reasons for the low nutritional value of sweet lupin seed meal are likely to be related to the chemical structure of the globulin proteins and their adverse effects on growth and nitrogen metabolism, and not to any known antinutritional factor or poor digestibility.
The aim of the present work was to study the nutritional utilization by rats of diets containing ... more The aim of the present work was to study the nutritional utilization by rats of diets containing chickpea or chickpea proteins purified from meal. Diets were formulated to contain the same amount of digestible energy (15.5 kJ/g) and protein (100 g/kg), and were supplemented with appropriate amounts of synthetic amino acids to target values. A lactalbumin-based diet was used as the control, and another diet containing defatted soybean as protein source was also included for practical comparisons. Feed intake was not affected by the inclusion of chickpea meal in the diet, but was lower in rats fed diets containing purified chickpea globulins. Performance (weight gains, gain:feed ratios) of rats fed chickpea meal or its globulins was lower than that of lactalbumin and soybean. Fecal and urinary N were increased in soybean and chickpea groups compared to lactalbumin. True N retention of rats fed diets containing lactalbumin (0.97) was higher than that of rats fed diets containing soybean, chickpea meal or its globulins (0.83–0.87). The plasma urea concentration of animals fed soybean- or chickpea-based diets increased in comparison with the lactalbumin group. The inclusion of chickpea insoluble residue in the control diet had no adverse effects on performance or N utilization. It is concluded that the reasons for the low nutritional value of chickpeas for growing animals are probably related mainly to the chemical structure of the globulin proteins and their adverse effects on growth and N metabolism.
An investigation into the anti-nutritive effect of feeding lupin seeds to male Hooded-Lister rats... more An investigation into the anti-nutritive effect of feeding lupin seeds to male Hooded-Lister rats, showed that lupin containing diets decreased the numbers of Escherichiu coli in the caecum, colon and the small intestine. This potentially beneficial lowering of E. coli populations in the gut may be due to the hindrance by the lupin fibre of the attachment of E. coli to gut receptors, since the effect also occurred in the small intestine where the lupin residue particles have not yet been degraded by bacterial fermentation. Accordingly, it may be possible to use lupin residue as a dietary supplement for reducing the number of potentially harmful E. coli in the monogastric gut.
The effects on the absorption of 65Zn by two varieties of raw faba bean (Vicia faba L., minor) or... more The effects on the absorption of 65Zn by two varieties of raw faba bean (Vicia faba L., minor) or seed components that may interfere with mineral metabolism in the gut, have been studied in growing rats. In bean diets all protein was supplied by the meals, and the fractions were tested by incorporating them in control diet at the same levels as they occur in the seeds. Absorption of 65Zn was also measured in rats fed dephytinized bean meal produced by including phytase in the diet. Rats were pair-fed diets supplemented with amino acids and minerals to target requirements and containing 40 mg Zn/kg diet. True absorption of Zn was 50-70% lower in rats fed diets containing both cultivars of faba bean meals than in those fed the control diet. Although soluble nonstarch polysaccharides caused a significant reduction in the absorption of Zn, this effect disappeared after the removal of phytate by demineralization. In contrast, despite its negligible content of phytate, the insoluble nonstarch polysaccharides in the cell wall fraction of the cotyledon accounted for most of the reduction in Zn absorption in rats fed the faba bean diets. Addition of phytic acid to the control diet significantly reduced the absorption of 65Zn but only from 44 to 36%. Moreover, the increase in the absorption of Zn was similarly small, from 21% to 29%, with the addition of phytase to the faba bean diet.(ABSTRACT TRUNCATED AT 250 WORDS)
Lactate is formed by many species of colonic bacteria, and can accumulate to high levels in the c... more Lactate is formed by many species of colonic bacteria, and can accumulate to high levels in the colons of inflammatory bowel disease subjects. Conversely, in healthy colons lactate is metabolized by lactate-utilizing species to the short-chain fatty acids butyrate and propionate, which are beneficial for the host. Here, we investigated the impact of continuous lactate infusions (up to 20 mM) at two pH values (6.5 and 5.5) on human colonic microbiota responsiveness and metabolic outputs. At pH 5.5 in particular, lactate tended to accumulate in tandem with decreases in butyrate and propionate and with corresponding changes in microbial composition. Moreover, microbial communities with low numbers of lactate-utilizing bacteria were inherently less stable and therefore more prone to lactate-induced perturbations. These investigations provide clear evidence of the important role these lactate utilizers may play in health maintenance. These should therefore be considered as potential new ...
Legume consumption has been reported to induce beneficial effects on obesity-associated metabolic... more Legume consumption has been reported to induce beneficial effects on obesity-associated metabolic disorders, but the underlying mechanisms have not been fully clarified. In the current work, pea (Pisum sativum L.) seed meal proteins (albumins, legumins and vicilins) were isolated, submitted to a simulated gastrointestinal digestion, and the effects of their hydrolysates (pea albumins hydrolysates (PAH), pea legumins hydrolysates (PLH) and pea vicilin hydrolysates (PVH), respectively) on 3T3-L1 murine pre-adipocytes were investigated. The pea vicilin hydrolysate (PVH), but not native pea vicilins, increased lipid accumulation during adipocyte differentiation. PVH also increased the mRNA expression levels of the adipocyte fatty acid-binding protein (aP2) and decreased that of pre-adipocyte factor-1 (Pref-1) (a pre-adipocyte marker gene), suggesting that PVH promotes adipocyte differentiation. Moreover, PVH induced adiponectin and insulin-responsive glucose transporter 4 (GLUT4) and st...
Overweight and obesity are regarded as world epidemics and are major risk factors for a number of... more Overweight and obesity are regarded as world epidemics and are major risk factors for a number of chronic diseases, including diabetes, cardiovascular diseases, and cancer. Two new highly palatable extruded mixes based on rice and pea (Pisum sativum) or kidney bean (Phaseolus vulgaris) meals were incorporated into normocaloric or obesogenic diets for rats at a low inclusion level (25%). Our purpose was to evaluate the effects of dietary incorporation of this new food ingredient on lipid profile. Organs (heart, liver, kidneys, spleen, stomach, small intestine, colon, cecum) and visceral fat relative weights were different (p < 0.01) from controls for animals fed the obesogenic diets and in rats fed extruded diets with respect to controls. Faecal excretion of bile acids was higher (p < 0.01) for rats fed extruded mixes compared with controls. The inclusion of extruded mixes replacing part of the casein in the control diet lowered liver cholesterol and triglycerides (p < 0.001...
Colorectal cancer is a worldwide disease with major clinical and economic impact, and its occurre... more Colorectal cancer is a worldwide disease with major clinical and economic impact, and its occurrence is determined by a variety of factors. In addition to its hereditary component, it is also known to be associated with various inflammatory processes, epigenetic alterations or modifications of the intestinal microbiota. The alimentary habits are crucial in the conformation of gut microbiota. The Mediterranean diet is widely recognized for its health benefits and has been associated with a lower risk of colon cancer. On the other hand, inflammation is a process commonly associated with cancer, and the intestinal microbiota interacts with the host to maintain normal function and health, particularly in processes of immunity and defense. Here, we are focusing in particular on two groups of substances (fibre, protein fractions) present in legumes whose mechanisms of action to prevent colon cancer or inflammation are likely to be mediated by the intestinal microbiota functional composition. 10 major chronic diseases including CRC (Kromhout, Spaaij, & de Goede, 2016; Song, Garrett, & Chan, 2015). In the current review, we are paying special attention to two groups of substances (fibre, protein fractions) present in legumes whose mechanisms of action to prevent
Desde el punto de vista de su uso en nutrición animal, pueden considerarse dos grupos de legumino... more Desde el punto de vista de su uso en nutrición animal, pueden considerarse dos grupos de leguminosas: leguminosas-forraje (p. ej. la alfalfa), usadas en alimentación de rumiantes y leguminosas-grano (p. ej. habas, guisantes y altramuces), usadas esencialmente para aves y cerdos, y en menor medida para rumiantes. Las leguminosas-grano pueden sustituir parcial o totalmente a fuentes de proteínas tradicionales, de origen animal, como carne, huesos o harina de pescado en los piensos para animales, y representan una alternativa para las harinas de soja y otras oleaginosas. Su contenido en proteína es variable pero elevado (25-45 g/100 g materia seca). No obstante, la presencia de metabolitos secundarios (inhibidores de proteasas, saponinas, glucósidos, lectinas, taninos, alcaloides), así como sus altos niveles de fibra (polisacáridos no amiláceos), ha restringido el uso de leguminosas-grano en la alimentación de animales monogástricos (aves y cerdos) y, mucho más en la de rumiantes. Sin embargo, en la actualidad se está incrementando el interés por el uso de estas materias primas como alimentos funcionales, sobre todo en nutrición humana, pero también en nutrición animal, entre otras razones, como consecuencia de la prohibición del uso de proteína animal (harinas de carne y hueso), que tuvo lugar en la Unión Europea tras la crisis de la Encefalopatía Espongiforme Bovina. Para establecer el valor nutritivo de las leguminosas ha de prestarse especial atención a su composición en nutrientes, su contenido en energía y la digestibilidad de los aminoácidos. En rumiantes, es necesario establecer la degradabilidad en el rumen tanto de la proteína como de los aminoácidos individuales y de los carbohidratos, aspectos que determinan la utilización de nutrientes en este grupo animal, pero sobre los que existe muy poca información. En la actualidad, la perspectiva respecto a los taninos en las dietas de rumiantes está cambiando considerándose que pueden ejercer un efecto beneficioso. La soja es la semilla más utilizada como fuente de proteína en nutrición animal, pero el hecho de que haya de importarse en su mayor parte y que sea modificada genéticamente suponen dificultades para su utilización en la actualidad en Europa. Razones de índole medioambiental, sanitarias y productivas resultan en un progresivo interés en el cultivo de leguminosas en la UE.
The effects of dietary supplementa tion with 2 recently developed feed additives on the compositi... more The effects of dietary supplementa tion with 2 recently developed feed additives on the composition of the mucosaassociated microbiota of the ileum were studied in growing broiler chickens. A total of 48 male 1-d-old broiler chickens of the Cobb 500 strain were distributed in 4 treatments with 2 repli cates of 6 birds each. The 2 additives tested were a di dfructose dianhydride-enriched caramel (FC) and the garlic derivative propyl propane thiosulfonate (PTS O). Dietary treatments were a control (commercial diet with no additive), INU (20 g inulin/kg diet), CAR (20 g FC/kg diet), and GAR (90 mgPTS-O/kg diet). As a result of this study, inulin supplementation resulted in lower (P < 0.05) and FC feeding resulted in higher (P < 0.05) Blautia coccoides/Eubacterium rectale log 10 number of copies respect to controls. Higher (P < 0.05) bifidobacteria log 10 number of copies with respect to the controls was determined in the ileal mucosa of birds fed the PTSO-supplemented diet. Denaturing gradient gel electrophoresis and PCR analysis on Bifidobacterium spp. revealed the presence of Bifidobacterium longum, Bifidobacterium pseudolongum, and Bifidobacterium pseudocatenulatum in samples from chickens fed the control and the PTS-O-supplemented diet. Bifidobacterium longum was exclusively found in poultry fed the control diet, whereas B. pseudocatenulatum was found only in poultry fed the PTSO-sup plemented diet. This study showed that both PTSO and FC were able to modulate the composition of the ileal mucosaassociated microbiota of growing broiler chickens. Finally, in addition to B. pseudolongum, the presence of B. longum and B. pseudocatenulatum, spe cies not previously described in intestinal samples of broilers, was also demonstrated.
Two experiments were carried out to study the effects of dietary supplementation with the garlic ... more Two experiments were carried out to study the effects of dietary supplementation with the garlic (Allium sativum)-derived product propyl propane thiosulfonate (PTS-O) on the intestinal log 10 number of copies of enteropathogens in broiler chickens, together with their intestinal morphology and growth performance. The additive had no significant effect on feed intake at any dose assayed. In experiment 1 (1 to 21 d of age), the BW of chickens fed on 45 mg of PTS-O/ kg of diet was higher (P < 0.01) than that of controls. Birds fed on diets containing 45 and 90 mg of PTS-O/ kg of diet had improved (P < 0.01) feed:gain ratios compared with the controls at 21 d of age. Ileal villus height, width and surface area, mucosal thickness, and muscular layer thickness were considerably greater (P < 0.01) than control values in chickens fed 90 mg of PTS-O/kg of diet. The Clostridium perfringens log 10 number of counts was not significantly affected at any dose assayed.
In vitroandin vivoexperiments were designed to evaluate the effectiveness of laboratory-made di-d... more In vitroandin vivoexperiments were designed to evaluate the effectiveness of laboratory-made di-d-fructose dianhydride (DFA)-enriched caramels. The DFA-enriched caramels were obtained fromd-fructose (FC),d-fructose and sucrose (FSC), ord-fructose and β-cyclodextrin (FCDC). In thein vitroexperiment, raftilose and all caramels increased (P<0.05)l-lactate concentration and decreased (P<0.05) pH. Total short-chain fatty acid concentration was higher (P<0.05) than controls in tubes containing raftilose, FSC, FCDC and commercial sucrose caramel (CSC). Raftilose, and all caramels tested except FSC and FC (1%), increased (P<0.01) lactobacilli log10number of copies compared with the non-additive control. FSC, FCDC and CSC increased (P<0.01) the bifidobacteria number of copies as compared with controls. All additives, except FCDC, decreased (P<0.01)Clostridium coccoides/Eubacterium rectalelog number of copies. Compared with controls, raftilose, FC and CSC led to lower (P<...
The effects oflupin (Lupinus angustifolius, cultivar 'Unicrop') seed meal or some of its fraction... more The effects oflupin (Lupinus angustifolius, cultivar 'Unicrop') seed meal or some of its fractions on the apparent and true absorption of minerals were studied in three experiments. Groups of four growing rats were pair-fed for 10 days (Experiments 1 and 2) orfor 14 days (Experiment 3) on diets which contained the same amounts of energy and protein, and supplemented with amino acids and minerals to target requirements. In Experiment 1, the apparent absorption of Ca, P and Zn was compared in animals fed a control (lactalbumin-based, LA) and a lupin meal (LM) diet. In addition, three lupin fractions were tested, two protein fractions (aqueous dialysed, soluble (ADS); aqueous dialysed, insoluble (ADI)) and a residue (LR) containing most of the material from the meal insoluble in water and phosphate-buffered saline (PBS). In Experiment 2, the true absorption of 6SZn was determined in three groups of rats given the following diets: LA (control), LM (lupin meal diet) and LR (diet containing the insoluble material). The inclusion of lupin protein fractions (ADS and ADI) in the diet had no significant effect on the apparent absorption of Ca, P or Zn. The apparent absorption of Ca was not affected by the inclusion in the diet of either lupin seed meal or its fractions but the apparent absorption of P was significantly lower in rats fed whole lupin meal diets (50.8%) than in those given the control diet (63.7%). The apparent absorption of Zn was also significantly lower in rats fed diets containing lupin meal (5.9%) or the insoluble residue (6,5%) compared with that of control animals (26.8%). The true absorption of 65Zn was decreased in animals fed on diets containing lupin meal (22.2%) or insoluble lupin residue (30.9%) in comparison with that of controls (41.9%), Growth was significantly retarded in rats given a lupin based diet with limited Zn supply (12 mg Zn kg-1) compared with those given a lupin diet containing 40 mg Zn kg-~ (Experiment 3). The results suggest that insoluble non-starch polysaccharides and phytate are mainly responsible for the lower availability of Zn in rats fed diets containing lupin meal as the only source of protein. Furthermore, growth can be
The effects on performance, digestibility, N utilization and plasma amino acid concentrations of ... more The effects on performance, digestibility, N utilization and plasma amino acid concentrations of dietary chickpea (Cicer arietinum, var. Kabuli) seed meal, globulin proteins or buffer-insoluble residue [starch / non-starch polysaccharides (NSP) / lignin] were studied in growing rats. Chickpea meal, defatted soybean meal, chickpea globulins and lactalbumin were each incorporated into diets as the sole source of dietary protein (100 g/kg). In addition, chickpea insoluble residue was included in a control diet in the same proportion found in the chickpea meal. Rats were killed while under halothane anesthesia after 10 d of consuming the diets, and ileal contents were washed out and freeze-dried for digestibility measurements. Weight gains and gain:feed ratios of rats fed chickpea diets for 10 d did not differ from those of rats fed defatted soybean but were significantly lower than those of rats given the control (lactalbumin) diet. However, ileal and fecal N digestibilities and N retention by rats fed the chickpea diet were significantly lower than those obtained with the lactalbumin or soybean diet. The inclusion of both chickpea meal or its globulin proteins in the diet significantly increased the amount of N excreted, primarily as urea, through the urine. However, although ileal N digestibility values for chickpea meal were significantly lower, those for its constituent globulins did not differ from control values. Urea levels in plasma in rats fed diets containing chickpea meal, globulins or soybean meal were significantly higher than in those fed lactalbumin. Furthermore, the concentrations of glycine, phenylalanine, histidine, arginine and ornithine in the plasma of rats fed chickpea meal, its globulins or defatted soybean were significantly higher, whereas those of threonine, leucine, lysine and tryptophan were significantly lower than lactalbumin-fed controls. The chickpea insoluble residue had no adverse effects on performance or N utilization by rats. We conclude that the low nutritional value of chickpea meal is likely to be due mainly to adverse effects of its globulin proteins on growth and N metabolism rather than to the action of any known antinutritional factor present in the diet.
Two experiments were carried out to study the effects of dietary supplementation with the garlic ... more Two experiments were carried out to study the effects of dietary supplementation with the garlic (Allium sativum)-derived product propyl propane thiosulfonate (PTS-O) on the intestinal log 10 number of copies of enteropathogens in broiler chickens, together with their intestinal morphology and growth performance. The additive had no significant effect on feed intake at any dose assayed. In experiment 1 (1 to 21 d of age), the BW of chickens fed on 45 mg of PTS-O/ kg of diet was higher (P < 0.01) than that of controls. Birds fed on diets containing 45 and 90 mg of PTS-O/ kg of diet had improved (P < 0.01) feed:gain ratios compared with the controls at 21 d of age. Ileal villus height, width and surface area, mucosal thickness, and muscular layer thickness were considerably greater (P < 0.01) than control values in chickens fed 90 mg of PTS-O/kg of diet. The Clostridium perfringens log 10 number of counts was not significantly affected at any dose assayed.
Castrated male Iberian (n = 12) and Landrace × Large White (n = 12) pigs were used to study histo... more Castrated male Iberian (n = 12) and Landrace × Large White (n = 12) pigs were used to study histological structure and enzymatic activity in the small intestine at 3 points of the productive cycle (BW = 15, 50, and 115 kg). Both strains were fed the same cereal-based diets (DE = 2,799 kcal•kg −1 , and CP = 15%) throughout the entire experimental period. Differences (P < 0.05) in histometrical variables (villus height, width and surface, crypt depth, villus height/ crypt depth relationship, mucosal thickness, muscular layer thickness, and number of goblet cells) were found among samples of small intestinal sections (duodenum, jejunum, and ileum) at the 3 productive stages studied. Also, differences (P < 0.05) in histometrical variables of small intestinal samples were found between Iberian and lean pigs at all productive stages, although these differences tended to disappear with age. Differences (P < 0.05) in enzymatic activities (lactase, sucrase, maltase, isomaltase, aminopeptidase, and alkaline phosphatase) of small intestinal samples were found between the different intestinal sections at all productive stages studied. Although differences (P < 0.05) in enzymatic activities of small intestinal samples were found between Iberian and lean pigs at all productive stages, values tended to equalize with pig age. We concluded that differences previously found in dietary nutritional utilization between Iberian and lean strains are likely not due to differences in intestinal absorption or hydrolytic capacity.
Cowpea (Vigna unguiculata) value as a functional food is still quite underexplored. Weaned Wistar... more Cowpea (Vigna unguiculata) value as a functional food is still quite underexplored. Weaned Wistar albino rats were fed one of four experimental equilibrated diets based on casein or cowpea meal as the only protein sources. RT-qPCR analysis showed that the replacement of casein for raw or treated cowpea meal lowered Escherichia/Shigella faecal counts and increased Lactobacilli, Bifidobacteria, Bacteroides, Clostridium leptum/Ruminococcus, and Faecalibacerium prausnitzii log 10 counts. Sequencing (Illumina) analysis confirmed these results and revealed that rats fed the cowpea-based diets presented higher proportions of potentially beneficial bacterial groups. Non-starch polysaccharides digestibility was associated with bacterial groups related with the control diet, while oligosaccharides digestibility was associated with cowpea diets. In conclusion, the inclusion of Vigna unguiculata in diets for rats, particularly in the untreated form, induced a healthier intestinal microbiota composition, which was mainly linked to the fermentation of oligosaccharides of the raffinose family.
Journal of the Science of Food and Agriculture, Jul 8, 2013
BACKGROUND: Legume seed proteins have to be chemically characterized in order to properly link th... more BACKGROUND: Legume seed proteins have to be chemically characterized in order to properly link their nutritional effects with their chemical structure. RESULTS: Vicilin and albumin fractions devoid of cross-contamination, as assessed by mass peptide fingerprinting analysis, were obtained from defatted pea (Pisum sativum cv. Bilbo) meal. The extracted protein fractions contained 56.7-67.7 g non-starch polysaccharides kg −1. The vicilin fraction was higher than legumins in arginine, isoleucine, leucine, phenylalanine and lysine. The most abundant amino acids in the albumin fraction were aspartic acid, glutamic acid, lysine and arginine, and the amounts of methionine were more than double than those in legumins and vicilins. The pea albumin fraction showed a clear enrichment of protease inhibitory activity when compared with the seed meal. In vitro digestibility values for pea proteins were 0.63 ± 0.04, 0.88 ± 0.04 and 0.41 ± 0.23 for legumins, vicilins and albumins respectively. CONCLUSION: Vicilin and albumin fractions devoid of cross-contamination with other proteins were obtained from pea seed meal. The vicilin fraction also contained low amounts of soluble non-starch polysaccharides and was enriched in isoleucine, leucine, phenylalanine and lysine. In vitro digestibility values for pea proteins were similar or even numerically higher than those for control proteins.
The effects of dietary sweet lupin (Lupinus angustifolius, Unicrop) seed meal or its insoluble fi... more The effects of dietary sweet lupin (Lupinus angustifolius, Unicrop) seed meal or its insoluble fiber (nonstarch polysaccharides + lignin) on performance, digestibility and nitrogen utilization in growing rats were studied in four experiments. Globulin proteins isolated from lupin, faba bean (Vicia faba L. minor) or soybean (Glycine max) were also incorporated into purified diets as replacements for lactalbumin (control) and the nutritional effects were evaluated. Isocaloric, legume-based diets supplemented with amino acids were used. Final weight gain, gain:feed ratios, nitrogen retention and net protein utilization of the animals fed whole lupin meal-based diets for 10 d were inferior to those of controls. In contrast, adding lupin insoluble fiber to a control diet produced no adverse effects. Ileal starch and apparent nitrogen digestibilities, and fecal digestibility of starch in lupin-fed rats were higher than those of controls, but fecal true nitrogen digestibility was lower. Replacement of lactalbumin with globulin proteins from lupin or faba bean depressed food intake and protein utilization, but only performance was affected by consumption of soybean globulins. Rats consuming lupin or faba bean globulins excreted significantly more nitrogen, particularly as urea through the urine. This did not occur in rats fed soybean globulins. Urea concentration in plasma was higher in rats fed diets containing lupin meal or legume globulins. The concentrations of urea, arginine and ornithine in plasma increased significantly compared with control values after 3 to 9 h of a lupin diet. After 9 h, plasma lysine was also decreased. We concluded that the main reasons for the low nutritional value of sweet lupin seed meal are likely to be related to the chemical structure of the globulin proteins and their adverse effects on growth and nitrogen metabolism, and not to any known antinutritional factor or poor digestibility.
The aim of the present work was to study the nutritional utilization by rats of diets containing ... more The aim of the present work was to study the nutritional utilization by rats of diets containing chickpea or chickpea proteins purified from meal. Diets were formulated to contain the same amount of digestible energy (15.5 kJ/g) and protein (100 g/kg), and were supplemented with appropriate amounts of synthetic amino acids to target values. A lactalbumin-based diet was used as the control, and another diet containing defatted soybean as protein source was also included for practical comparisons. Feed intake was not affected by the inclusion of chickpea meal in the diet, but was lower in rats fed diets containing purified chickpea globulins. Performance (weight gains, gain:feed ratios) of rats fed chickpea meal or its globulins was lower than that of lactalbumin and soybean. Fecal and urinary N were increased in soybean and chickpea groups compared to lactalbumin. True N retention of rats fed diets containing lactalbumin (0.97) was higher than that of rats fed diets containing soybean, chickpea meal or its globulins (0.83–0.87). The plasma urea concentration of animals fed soybean- or chickpea-based diets increased in comparison with the lactalbumin group. The inclusion of chickpea insoluble residue in the control diet had no adverse effects on performance or N utilization. It is concluded that the reasons for the low nutritional value of chickpeas for growing animals are probably related mainly to the chemical structure of the globulin proteins and their adverse effects on growth and N metabolism.
An investigation into the anti-nutritive effect of feeding lupin seeds to male Hooded-Lister rats... more An investigation into the anti-nutritive effect of feeding lupin seeds to male Hooded-Lister rats, showed that lupin containing diets decreased the numbers of Escherichiu coli in the caecum, colon and the small intestine. This potentially beneficial lowering of E. coli populations in the gut may be due to the hindrance by the lupin fibre of the attachment of E. coli to gut receptors, since the effect also occurred in the small intestine where the lupin residue particles have not yet been degraded by bacterial fermentation. Accordingly, it may be possible to use lupin residue as a dietary supplement for reducing the number of potentially harmful E. coli in the monogastric gut.
The effects on the absorption of 65Zn by two varieties of raw faba bean (Vicia faba L., minor) or... more The effects on the absorption of 65Zn by two varieties of raw faba bean (Vicia faba L., minor) or seed components that may interfere with mineral metabolism in the gut, have been studied in growing rats. In bean diets all protein was supplied by the meals, and the fractions were tested by incorporating them in control diet at the same levels as they occur in the seeds. Absorption of 65Zn was also measured in rats fed dephytinized bean meal produced by including phytase in the diet. Rats were pair-fed diets supplemented with amino acids and minerals to target requirements and containing 40 mg Zn/kg diet. True absorption of Zn was 50-70% lower in rats fed diets containing both cultivars of faba bean meals than in those fed the control diet. Although soluble nonstarch polysaccharides caused a significant reduction in the absorption of Zn, this effect disappeared after the removal of phytate by demineralization. In contrast, despite its negligible content of phytate, the insoluble nonstarch polysaccharides in the cell wall fraction of the cotyledon accounted for most of the reduction in Zn absorption in rats fed the faba bean diets. Addition of phytic acid to the control diet significantly reduced the absorption of 65Zn but only from 44 to 36%. Moreover, the increase in the absorption of Zn was similarly small, from 21% to 29%, with the addition of phytase to the faba bean diet.(ABSTRACT TRUNCATED AT 250 WORDS)
Lactate is formed by many species of colonic bacteria, and can accumulate to high levels in the c... more Lactate is formed by many species of colonic bacteria, and can accumulate to high levels in the colons of inflammatory bowel disease subjects. Conversely, in healthy colons lactate is metabolized by lactate-utilizing species to the short-chain fatty acids butyrate and propionate, which are beneficial for the host. Here, we investigated the impact of continuous lactate infusions (up to 20 mM) at two pH values (6.5 and 5.5) on human colonic microbiota responsiveness and metabolic outputs. At pH 5.5 in particular, lactate tended to accumulate in tandem with decreases in butyrate and propionate and with corresponding changes in microbial composition. Moreover, microbial communities with low numbers of lactate-utilizing bacteria were inherently less stable and therefore more prone to lactate-induced perturbations. These investigations provide clear evidence of the important role these lactate utilizers may play in health maintenance. These should therefore be considered as potential new ...
Legume consumption has been reported to induce beneficial effects on obesity-associated metabolic... more Legume consumption has been reported to induce beneficial effects on obesity-associated metabolic disorders, but the underlying mechanisms have not been fully clarified. In the current work, pea (Pisum sativum L.) seed meal proteins (albumins, legumins and vicilins) were isolated, submitted to a simulated gastrointestinal digestion, and the effects of their hydrolysates (pea albumins hydrolysates (PAH), pea legumins hydrolysates (PLH) and pea vicilin hydrolysates (PVH), respectively) on 3T3-L1 murine pre-adipocytes were investigated. The pea vicilin hydrolysate (PVH), but not native pea vicilins, increased lipid accumulation during adipocyte differentiation. PVH also increased the mRNA expression levels of the adipocyte fatty acid-binding protein (aP2) and decreased that of pre-adipocyte factor-1 (Pref-1) (a pre-adipocyte marker gene), suggesting that PVH promotes adipocyte differentiation. Moreover, PVH induced adiponectin and insulin-responsive glucose transporter 4 (GLUT4) and st...
Overweight and obesity are regarded as world epidemics and are major risk factors for a number of... more Overweight and obesity are regarded as world epidemics and are major risk factors for a number of chronic diseases, including diabetes, cardiovascular diseases, and cancer. Two new highly palatable extruded mixes based on rice and pea (Pisum sativum) or kidney bean (Phaseolus vulgaris) meals were incorporated into normocaloric or obesogenic diets for rats at a low inclusion level (25%). Our purpose was to evaluate the effects of dietary incorporation of this new food ingredient on lipid profile. Organs (heart, liver, kidneys, spleen, stomach, small intestine, colon, cecum) and visceral fat relative weights were different (p < 0.01) from controls for animals fed the obesogenic diets and in rats fed extruded diets with respect to controls. Faecal excretion of bile acids was higher (p < 0.01) for rats fed extruded mixes compared with controls. The inclusion of extruded mixes replacing part of the casein in the control diet lowered liver cholesterol and triglycerides (p < 0.001...
Colorectal cancer is a worldwide disease with major clinical and economic impact, and its occurre... more Colorectal cancer is a worldwide disease with major clinical and economic impact, and its occurrence is determined by a variety of factors. In addition to its hereditary component, it is also known to be associated with various inflammatory processes, epigenetic alterations or modifications of the intestinal microbiota. The alimentary habits are crucial in the conformation of gut microbiota. The Mediterranean diet is widely recognized for its health benefits and has been associated with a lower risk of colon cancer. On the other hand, inflammation is a process commonly associated with cancer, and the intestinal microbiota interacts with the host to maintain normal function and health, particularly in processes of immunity and defense. Here, we are focusing in particular on two groups of substances (fibre, protein fractions) present in legumes whose mechanisms of action to prevent colon cancer or inflammation are likely to be mediated by the intestinal microbiota functional composition. 10 major chronic diseases including CRC (Kromhout, Spaaij, & de Goede, 2016; Song, Garrett, & Chan, 2015). In the current review, we are paying special attention to two groups of substances (fibre, protein fractions) present in legumes whose mechanisms of action to prevent
Desde el punto de vista de su uso en nutrición animal, pueden considerarse dos grupos de legumino... more Desde el punto de vista de su uso en nutrición animal, pueden considerarse dos grupos de leguminosas: leguminosas-forraje (p. ej. la alfalfa), usadas en alimentación de rumiantes y leguminosas-grano (p. ej. habas, guisantes y altramuces), usadas esencialmente para aves y cerdos, y en menor medida para rumiantes. Las leguminosas-grano pueden sustituir parcial o totalmente a fuentes de proteínas tradicionales, de origen animal, como carne, huesos o harina de pescado en los piensos para animales, y representan una alternativa para las harinas de soja y otras oleaginosas. Su contenido en proteína es variable pero elevado (25-45 g/100 g materia seca). No obstante, la presencia de metabolitos secundarios (inhibidores de proteasas, saponinas, glucósidos, lectinas, taninos, alcaloides), así como sus altos niveles de fibra (polisacáridos no amiláceos), ha restringido el uso de leguminosas-grano en la alimentación de animales monogástricos (aves y cerdos) y, mucho más en la de rumiantes. Sin embargo, en la actualidad se está incrementando el interés por el uso de estas materias primas como alimentos funcionales, sobre todo en nutrición humana, pero también en nutrición animal, entre otras razones, como consecuencia de la prohibición del uso de proteína animal (harinas de carne y hueso), que tuvo lugar en la Unión Europea tras la crisis de la Encefalopatía Espongiforme Bovina. Para establecer el valor nutritivo de las leguminosas ha de prestarse especial atención a su composición en nutrientes, su contenido en energía y la digestibilidad de los aminoácidos. En rumiantes, es necesario establecer la degradabilidad en el rumen tanto de la proteína como de los aminoácidos individuales y de los carbohidratos, aspectos que determinan la utilización de nutrientes en este grupo animal, pero sobre los que existe muy poca información. En la actualidad, la perspectiva respecto a los taninos en las dietas de rumiantes está cambiando considerándose que pueden ejercer un efecto beneficioso. La soja es la semilla más utilizada como fuente de proteína en nutrición animal, pero el hecho de que haya de importarse en su mayor parte y que sea modificada genéticamente suponen dificultades para su utilización en la actualidad en Europa. Razones de índole medioambiental, sanitarias y productivas resultan en un progresivo interés en el cultivo de leguminosas en la UE.
The effects of dietary supplementa tion with 2 recently developed feed additives on the compositi... more The effects of dietary supplementa tion with 2 recently developed feed additives on the composition of the mucosaassociated microbiota of the ileum were studied in growing broiler chickens. A total of 48 male 1-d-old broiler chickens of the Cobb 500 strain were distributed in 4 treatments with 2 repli cates of 6 birds each. The 2 additives tested were a di dfructose dianhydride-enriched caramel (FC) and the garlic derivative propyl propane thiosulfonate (PTS O). Dietary treatments were a control (commercial diet with no additive), INU (20 g inulin/kg diet), CAR (20 g FC/kg diet), and GAR (90 mgPTS-O/kg diet). As a result of this study, inulin supplementation resulted in lower (P < 0.05) and FC feeding resulted in higher (P < 0.05) Blautia coccoides/Eubacterium rectale log 10 number of copies respect to controls. Higher (P < 0.05) bifidobacteria log 10 number of copies with respect to the controls was determined in the ileal mucosa of birds fed the PTSO-supplemented diet. Denaturing gradient gel electrophoresis and PCR analysis on Bifidobacterium spp. revealed the presence of Bifidobacterium longum, Bifidobacterium pseudolongum, and Bifidobacterium pseudocatenulatum in samples from chickens fed the control and the PTS-O-supplemented diet. Bifidobacterium longum was exclusively found in poultry fed the control diet, whereas B. pseudocatenulatum was found only in poultry fed the PTSO-sup plemented diet. This study showed that both PTSO and FC were able to modulate the composition of the ileal mucosaassociated microbiota of growing broiler chickens. Finally, in addition to B. pseudolongum, the presence of B. longum and B. pseudocatenulatum, spe cies not previously described in intestinal samples of broilers, was also demonstrated.
Two experiments were carried out to study the effects of dietary supplementation with the garlic ... more Two experiments were carried out to study the effects of dietary supplementation with the garlic (Allium sativum)-derived product propyl propane thiosulfonate (PTS-O) on the intestinal log 10 number of copies of enteropathogens in broiler chickens, together with their intestinal morphology and growth performance. The additive had no significant effect on feed intake at any dose assayed. In experiment 1 (1 to 21 d of age), the BW of chickens fed on 45 mg of PTS-O/ kg of diet was higher (P < 0.01) than that of controls. Birds fed on diets containing 45 and 90 mg of PTS-O/ kg of diet had improved (P < 0.01) feed:gain ratios compared with the controls at 21 d of age. Ileal villus height, width and surface area, mucosal thickness, and muscular layer thickness were considerably greater (P < 0.01) than control values in chickens fed 90 mg of PTS-O/kg of diet. The Clostridium perfringens log 10 number of counts was not significantly affected at any dose assayed.
In vitroandin vivoexperiments were designed to evaluate the effectiveness of laboratory-made di-d... more In vitroandin vivoexperiments were designed to evaluate the effectiveness of laboratory-made di-d-fructose dianhydride (DFA)-enriched caramels. The DFA-enriched caramels were obtained fromd-fructose (FC),d-fructose and sucrose (FSC), ord-fructose and β-cyclodextrin (FCDC). In thein vitroexperiment, raftilose and all caramels increased (P<0.05)l-lactate concentration and decreased (P<0.05) pH. Total short-chain fatty acid concentration was higher (P<0.05) than controls in tubes containing raftilose, FSC, FCDC and commercial sucrose caramel (CSC). Raftilose, and all caramels tested except FSC and FC (1%), increased (P<0.01) lactobacilli log10number of copies compared with the non-additive control. FSC, FCDC and CSC increased (P<0.01) the bifidobacteria number of copies as compared with controls. All additives, except FCDC, decreased (P<0.01)Clostridium coccoides/Eubacterium rectalelog number of copies. Compared with controls, raftilose, FC and CSC led to lower (P<...
The effects oflupin (Lupinus angustifolius, cultivar 'Unicrop') seed meal or some of its fraction... more The effects oflupin (Lupinus angustifolius, cultivar 'Unicrop') seed meal or some of its fractions on the apparent and true absorption of minerals were studied in three experiments. Groups of four growing rats were pair-fed for 10 days (Experiments 1 and 2) orfor 14 days (Experiment 3) on diets which contained the same amounts of energy and protein, and supplemented with amino acids and minerals to target requirements. In Experiment 1, the apparent absorption of Ca, P and Zn was compared in animals fed a control (lactalbumin-based, LA) and a lupin meal (LM) diet. In addition, three lupin fractions were tested, two protein fractions (aqueous dialysed, soluble (ADS); aqueous dialysed, insoluble (ADI)) and a residue (LR) containing most of the material from the meal insoluble in water and phosphate-buffered saline (PBS). In Experiment 2, the true absorption of 6SZn was determined in three groups of rats given the following diets: LA (control), LM (lupin meal diet) and LR (diet containing the insoluble material). The inclusion of lupin protein fractions (ADS and ADI) in the diet had no significant effect on the apparent absorption of Ca, P or Zn. The apparent absorption of Ca was not affected by the inclusion in the diet of either lupin seed meal or its fractions but the apparent absorption of P was significantly lower in rats fed whole lupin meal diets (50.8%) than in those given the control diet (63.7%). The apparent absorption of Zn was also significantly lower in rats fed diets containing lupin meal (5.9%) or the insoluble residue (6,5%) compared with that of control animals (26.8%). The true absorption of 65Zn was decreased in animals fed on diets containing lupin meal (22.2%) or insoluble lupin residue (30.9%) in comparison with that of controls (41.9%), Growth was significantly retarded in rats given a lupin based diet with limited Zn supply (12 mg Zn kg-1) compared with those given a lupin diet containing 40 mg Zn kg-~ (Experiment 3). The results suggest that insoluble non-starch polysaccharides and phytate are mainly responsible for the lower availability of Zn in rats fed diets containing lupin meal as the only source of protein. Furthermore, growth can be
The effects on performance, digestibility, N utilization and plasma amino acid concentrations of ... more The effects on performance, digestibility, N utilization and plasma amino acid concentrations of dietary chickpea (Cicer arietinum, var. Kabuli) seed meal, globulin proteins or buffer-insoluble residue [starch / non-starch polysaccharides (NSP) / lignin] were studied in growing rats. Chickpea meal, defatted soybean meal, chickpea globulins and lactalbumin were each incorporated into diets as the sole source of dietary protein (100 g/kg). In addition, chickpea insoluble residue was included in a control diet in the same proportion found in the chickpea meal. Rats were killed while under halothane anesthesia after 10 d of consuming the diets, and ileal contents were washed out and freeze-dried for digestibility measurements. Weight gains and gain:feed ratios of rats fed chickpea diets for 10 d did not differ from those of rats fed defatted soybean but were significantly lower than those of rats given the control (lactalbumin) diet. However, ileal and fecal N digestibilities and N retention by rats fed the chickpea diet were significantly lower than those obtained with the lactalbumin or soybean diet. The inclusion of both chickpea meal or its globulin proteins in the diet significantly increased the amount of N excreted, primarily as urea, through the urine. However, although ileal N digestibility values for chickpea meal were significantly lower, those for its constituent globulins did not differ from control values. Urea levels in plasma in rats fed diets containing chickpea meal, globulins or soybean meal were significantly higher than in those fed lactalbumin. Furthermore, the concentrations of glycine, phenylalanine, histidine, arginine and ornithine in the plasma of rats fed chickpea meal, its globulins or defatted soybean were significantly higher, whereas those of threonine, leucine, lysine and tryptophan were significantly lower than lactalbumin-fed controls. The chickpea insoluble residue had no adverse effects on performance or N utilization by rats. We conclude that the low nutritional value of chickpea meal is likely to be due mainly to adverse effects of its globulin proteins on growth and N metabolism rather than to the action of any known antinutritional factor present in the diet.
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