Dietary Intake, Nutritional Adequacy, and Food Sources of Protein and Relationships with Personal and Family Factors in Spanish Children Aged One to <10 Years: Findings of the EsNuPI Study.

Diet in the first years of life is an important factor in growth and development. Dietary protein is a critical macronutrient that provides both essential and nonessential amino acids required for sustaining all body functions and procedures, providing the structural basis to maintain life and healthy development and growth in children. In this study, our aim was to describe the total protein intake, type and food sources of protein, the adequacy to the Population Reference Intake (PRI) for protein by the European Food Safety Authority (EFSA), and the Recommended Dietary Allowance (RDA) by the Institute of Medicine (IoM). Furthermore, we analyzed whether the consumption of dairy products (including regular milk, dairy products, or adapted milk formulas) is associated with nutrient adequacy and the contribution of protein to diet and whole dietary profile in the two cohorts of the EsNuPI (in English, Nutritional Study in the Spanish Pediatric Population) study; one cohort was representative of the Spanish population from one to <10 years old (n = 707) (Spanish reference cohort, SRS) who reported consuming all kinds of milk and one was a cohort of the same age who reported consuming adapted milk over the last year (including follow-on formula, growing up milk, toddler's milk, and enriched and fortified milks) (n = 741) (adapted milk consumers cohort, AMS). The children of both cohorts had a high contribution from protein to total energy intake (16.79% SRS and 15.63% AMS) and a high total protein intake (60.89 g/day SRS and 53.43 g/day AMS). We observed that protein intake in Spanish children aged one to <10 years old was above the European and international recommendations, as well as the recommended percentages for energy intakes. The main protein sources were milk and dairy products (28% SRS and 29% AMS) and meat and meat products (27% SRS and 26% AMS), followed by cereals (16% SRS and 15% AMS), fish and shellfish (8% in both cohorts), eggs (5% SRS and 6% AMS), and legumes (4% in both cohorts). In our study population, protein intake was mainly from an animal origin (meat and meat products, milk and dairy products, fish and shellfish, and eggs) rather than from a plant origin (cereals and legumes). Future studies should investigate the long-term effect of dietary protein in early childhood on growth and body composition, and whether high protein intake affects health later in life.

Clustering of Dietary Patterns and Lifestyles Among Spanish Children in the EsNuPI Study †.

Dietary patterns (DPs) are known to be tied to lifestyle behaviors. Understanding DPs and their relationships with lifestyle factors can help to prevent children from engaging in unhealthy dietary practices. We aimed to describe DPs in Spanish children aged 1 to <10 years and to examine their associations with sociodemographic and lifestyle variables. The consumption of toddler and young children milk formulas, enriched and fortified milk within the Spanish pediatric population is increasing, and there is a lack of evidence whether the consumption of this type of milk is causing an impact on nutrient intakes and if they are helping to reach the nutrient recommendations. Within the Nutritional Study in the Spanish Pediatric Population (EsNuPI), we considered two study cohorts and three different age groups in three year-intervals in each of them. The study cohort included 740 children in a representative sample of the urban non-vegan Spanish population and 772 children in a convenience cohort of adapted milk consumers (AMS) (including follow-on formula, toddler's milk, growing up milk, and fortified and enriched milks) who provided information about sociodemographics, lifestyle, and dietary habits; a food frequency questionnaire was used for the latter. Principal component analysis was performed to identify DPs from 18 food groups. Food groups and sociodemographic/lifestyle variables were combined through a hierarchical cluster algorithm. Three DPs predominated in every age group and study sample: a palatable energy-dense food dietary pattern, and two Mediterranean-like DPs. However, children from the AMS showed a predominant dietary pattern markedly related to the Mediterranean diet, with high consumption of cereals, fruits and vegetables, as well as milk and dairy products. The age of children and certain lifestyle factors, namely level of physical activity, parental education, and household income, correlated closely with the dietary clusters. Thus, the findings provide insight into designing lifestyle interventions that could reverse the appearance of unhealthy DPs in the Spanish child population.

Dietary Intake, Nutritional Adequacy and Food Sources of Total Fat and Fatty Acids, and Relationships with Personal and Family Factors in Spanish Children Aged One to <10 Years: Results of the EsNuPI Study.

We aimed to determine the usual intake of total fat, fatty acids (FAs), and their main food sources in a representative cohort of the Spanish pediatric population aged 1 to <10 years (n = 707) who consumed all types of milk and an age-matched cohort who consumed adapted milk over the last year (including follow-on formula, toddler's milk, growing-up milk, and fortified and enriched milks) (n = 741) who were participants in the EsNuPI study (in English, Nutritional Study in the Spanish Pediatric Population). Dietary intake, measured through two 24 h dietary recalls, was compared to the European Food Safety Authority (EFSA) and the Food and Agriculture Organization of the United Nations (UN-FAO) recommendations. Both cohorts showed a high intake of saturated fatty acids (SFAs), according to FAO recommendations, as there are no numerical recommendations for SFAs at EFSA. Also, low intake of essential fatty acids (EFAs; linoleic acid (LA) and α-linolenic acid (ALA)) and long-chain polyunsaturated fatty acids (LC-PUFA) of the n-3 series, mainly docosahexaenoic acid (DHA) were observed according to EFSA and FAO recommendations. The three main sources of total fat and different FAs were milk and dairy products, oils and fats, and meat and meat products. The consumption of adapted milk was one of the main factors associated with better adherence to the nutritional recommendations of total fat, SFAs, EFAs, PUFAs; and resulted as the main factor associated with better adherence to n-3 fatty acids intake recommendations. Knowledge of the dietary intake and food sources of total fat and FAs in children could help in designing and promoting effective and practical age-targeted guidelines to promote the consumption of EFA- and n-3 PUFA-rich foods in this stage of life.

Usual Dietary Intake, Nutritional Adequacy and Food Sources of Calcium, Phosphorus, Magnesium and Vitamin D of Spanish Children Aged One to <10 Years. Findings from the EsNuPI Study.

Bone problems in the population begin to be establish in childhood. The present study aims to assess the usual calcium, phosphorus, magnesium, and vitamin D intakes, along with the food sources of these nutrients, in Spanish children participating in the EsNuPI (Estudio Nutricional en Población Infantil Española) study. Two 24 h dietary recalls were applied to 1448 children (1 to <10 years) divided into two sub-samples: one reference sample (RS) of the general population [n = 707] and another sample which exclusively included children consuming enriched or fortified milks, here called "adapted milks" (AMS) [n = 741]. Estimation of the usual intake shows that nutrient intake increased with age for all nutrients except vitamin D. Using as reference the Dietary Reference Values from the European Food Safety Authority (EFSA), calcium and magnesium intakes were found to be below the average requirement (AR) and adequate intake (AI), respectively, in a considerable percentage of children. Furthermore, phosphorus exceeded the AI in 100% of individuals and vitamin D was lower than the AI in almost all children studied. The results were very similar when considering only plausible reporters. When analyzing the food sources of the nutrients studied, milk and dairy products contributed the most to calcium, phosphorus, magnesium, and vitamin D. Other sources of calcium were cereals and vegetables; for phosphorus: meat, meat products, and cereals; for magnesium: cereals and fruits; and, for vitamin D: fish and eggs. These results highlight the desirability of improving the intake concerning these nutrients, which are involved in bone and metabolic health in children. The AMS group appeared to contribute better to the adequacy of those nutrients than the RS group, but both still need further improvement. Of special interest are the results of vitamin D intakes, which were significantly higher in the AMS group (although still below the AI), independent of age.

Dietary and Lifestyle Patterns in the Spanish Pediatric Population (One to <10 Years Old): Design, Protocol, and Methodology of the EsNuPI Study.

The interest in a healthy diet and lifestyle during the early stages of life increased, pointing out its role in the development of noncommunicable chronic diseases throughout adult life. Dietary habits and dietary patterns begin to be established in early childhood and persist during adulthood. Therefore, the EsNuPI ("Nutritional Study in Spanish Pediatric Population") study aims to depict the dietary patterns, physical activity, and sedentary behaviors in Spanish children aged from one to <10 years old. This prospective, cross-sectional, observational study recruited a total of 1514 children from Spanish cities with >50,000 inhabitants, stratified by Nielsen areas. Participants were involved in one face-to-face survey, followed by a telephone survey after at least one week. Information about dietary intake and habits was obtained using a quantitative food frequency questionnaire and two 24-h dietary recalls. Physical activity and sedentary behaviors were registered using a specific questionnaire based on a seven-day record. Data were processed and stratified by categorical variables to be statistically analyzed in order to meet the study objectives. This study is the first of its kind in a Spanish reference population of this age range and the first to evaluate whether the consumption of adapted milk formulas and dairy products is associated with healthier dietary patterns and better diet quality and lifestyles in this group.

Factors Predicting the Response to a Vitamin D-Fortified Milk in Healthy Postmenopausal Women.

BACKGROUND: Milk products fortified with vitamin D may constitute an alternative to pharmacological supplements for reaching the optimal levels of serum 25-hydroxyvitamin D [25(OH)D]. Our aim was to analyze the response of serum 25(OH)D and its predictive factors in postmenopausal healthy women after a dietary intervention with a milk fortified with vitamin D and calcium. METHODS: We designed a prospective study including 305 healthy postmenopausal women who consumed a fortified milk with calcium (900 mg/500 mL) and vitamin D3 (600 IU/500 mL) daily for 24 months. RESULTS: The 25(OH)D concentrations at 24 months were correlated to weight, to body mass index, to the percentage of fat, triglycerides and to baseline 25(OH)D levels. We found significant differences in the levels of 25(OH)D at 24 months according to baseline 25(OH)D levels (p < 0.001) and body mass index (p = 0.019) expressed at quartiles. Multivariate analysis showed an association between levels of 25(OH)D after the intervention and at baseline 25(OH)D (Beta = 0.47, p < 0.001) and percentage of body fat (Beta = -0.227, p = 0.049), regardless of the body mass index. CONCLUSIONS: In healthy postmenopausal women, the improvement in 25(OH)D after an intervention with a fortified milk for 24 months depends mainly on the baseline levels of serum 25(OH)D and on the percentage of body fat.

Effects of Daily Intake of Calcium and Vitamin D-Enriched Milk in Healthy Postmenopausal Women: A Randomized, Controlled, Double-Blind Nutritional Study.

OBJECTIVE: To determine the effect of the daily intake of calcium and vitamin D-enriched milk (with or without fructooligosaccharides [FOS]) on vitamin D, bone metabolism, and cardiovascular risk factors. MATERIALS AND METHODS: Two-year randomized controlled study, including 500 healthy postmenopausal women, assigned to 500 mL/day of skimmed milk to one of three groups: Low-dose (L): (120 mg/100 mL calcium, vitamin D3 30 UI/100 mL), group A: calcium and vitamin D (180 mg/100 mL and 120 UI/100 mL), and group B: calcium and vitamin D (180 mg/100 mL and 120 UI/100 mL) and FOS (5 g/L). We evaluated serum 25(OH)D, bone mineral density (BMD) by Dual Energy X-ray Absorptiometry, and biochemical data of glucose and lipid metabolism. RESULTS: After 24 months, vitamin D concentrations did not change in the control group, but increased in group A and group B, p < 0.001. We observed an increase in femoral neck BMD and an improvement in fasting plasma glucose, HbA1c, total cholesterol, low-density lipoprotein cholesterol, and apolipoprotein B 100. CONCLUSIONS: Daily intake of milk enriched with calcium and vitamin D in postmenopausal healthy women induces a significant improvement in vitamin D status, a significant increase in BMD at femoral neck, and also favorable effects on glucose and lipid profile.

Omega-3 LCPUFA supplement: a nutritional strategy to prevent maternal and neonatal oxidative stress.

There is controversy about fish-oil supplementation and oxidative damage. This ambiguity should be explored to elucidate its role as modulator of oxidative stress, especially during gestation and postnatal life. This is the objective of this study. One hundred ten pregnant women were divided in two groups: control group CT (400 mL/day of the control dairy drink); supplemented group FO (400 mL/day of the fish oil-enriched dairy drink (±400-mg EPA-DHA/day)). Different biomarkers of oxidative damage were determined in the mother's at enrolment, at delivery and at 2.5 and 4 months postpartum and newborns at delivery and at 2.5 months postpartum. Omega-3 LC-PUFA supplementation during pregnancy and lactation decreased plasma hydroperoxides especially in newborn at delivery (P = 0.001) and 2.5 months (P = 0.006), increased superoxide dismutase (SOD) and catalase (CAT) in mothers at delivery (P = 0.024 (SOD)) and after 2.5 months (P = 0.040 (CAT)) and in newborns at 2.5 months (P = 0.035 (SOD); P = 0.021 (CAT)). Also, supplementation increased α-tocoferol in mothers at 2.5 months (P = 0.030) and in umbilical cord artery (P = 0.039). Higher levels of CoQ10 were found in mothers at delivery (P = 0.039) as well as in umbilical cord vein (P = 0.024) and artery (P = 0.036). Our supplementation prevents the oxidative stress in the mother and neonate during the first months of postnatal life, being a potential preventive nutritional strategy to prevent functional alterations associated with oxidative stress that have an important repercussion for the neonate development in the early postnatal life.

Effects of Maternal Ω-3 Supplementation on Fatty Acids and on Visual and Cognitive Development.

OBJECTIVES: The aim of the present study was to elucidate whether a dairy drink enriched with ω-3 long-chain polyunsaturated fatty acid (LC-PUFA) could have an impact on the lipid profile of the mother and the newborn, and also whether this intervention could affect the newborns' visual and cognitive development. METHODS: A total of 110 pregnant women were randomly assigned to one of the following intervention groups: control group (n = 54), taking 400 mL/day of the control dairy drink, and supplemented group (fish oil [FO]) (n = 56), taking 400 mL/day of the fish oil-enriched dairy drink (including ∼400 mg eicosapentaenoic acid-docosahexaenoic acid [DHA]/day). During the study, the mothers' diets were supervised by a nutritionist to encourage compliance with present recommendations of FA intake. Blood fatty acid profiles were determined in the mother's (at enrollment, at delivery, and at 2.5 and 4 months) and newborn (at delivery and at 2.5 months) placenta and breast milk (colostrum and at 1, 2, and 4 months). Pattern reversal visual evoked potentials (VEPs) (at 2.5 and 7.5 months) and Bayley test (at 12 months) were recorded. RESULTS: DHA percentage was higher in plasma, erythrocyte membranes, and breast milk samples from the FO group. The ratio of nervonic acid was also higher in plasma and erythrocyte lipids of the mother and newborn's blood samples from the FO group. No differences were observed in the Bayley test. No differences were observed in VEPs between both groups. We observed a shorter latency, however, in the lower visual angle (7.5') in the boys of the supplemented group. CONCLUSIONS: Omega-3 LC-PUFA dietary supplement during pregnancy and lactation influenced the mother and newborn's fatty acid profile and nervonic acid content but did not show effects on visual and cognitive/psychomotor development.

A genetic variant of PPARA modulates cardiovascular risk biomarkers after milk consumption.

OBJECTIVE: The association of dairy food consumption with the risk for developing cardiovascular disease (CVD) has been investigated in many studies, but results often have been contradictory. The aim of the present study was to determine whether genetic polymorphisms are associated with interindividual variation in the response of CVD risk biomarker values after milk consumption. METHODS: Fourteen single nucleotide polymorphisms (SNPs) in nine genes related to lipid metabolism were examined in 161 volunteers randomly allocated to consume either 500 mL/d of skimmed (S) or semi-skimmed (SS) milk for 1 year in addition to their usual diets. Total cholesterol/high-density lipoprotein cholesterol (TC/HDL-C) and low-density lipoprotein/HDL-C ratios were used as biomarkers of CVD risk. Three-way repeated-measures analysis of variance was used to examine the effect of time, treatment (S or SS), and genotype on these biomarkers. RESULTS: A TT genotype for the proliferator-activated receptor alpha polymorphism (PPARA rs135549 SNP) was significantly associated with a reduction in the TC/HDL and LDL/HDL ratios after 12 mo of S milk intake (mean reduction -0.29, 95% confidence interval [CI], -0.63 to 0.05; P = 0.0015 and -0.31, 95% CI, -0.58 to -0.03; P = 0.0005, respectively). However, no differences were observed after consuming either S or SS milk in the C allele carriers. CONCLUSIONS: Saturated fatty acid consumption has long been linked to an increased risk for CVD; indeed, the consumption of saturated fat-free products is recommended as a means of reducing this risk. However, the present results suggest that many individuals might not benefit from such general recommendations. Genetic analysis of PPARA rs135549 might help identify those individuals who are more likely to benefit from reducing the saturated fatty acid content of their diet.

Human milk probiotic Lactobacillus fermentum CECT5716 reduces the incidence of gastrointestinal and upper respiratory tract infections in infants.

OBJECTIVES: The aim of the study was to examine the effects of a follow-on formula containing Lactobacillus fermentum CECT5716 (L. fermentum) on the incidence of infections in infants between the ages of 6 and 12 months. PATIENTS AND METHODS: A randomized double-blinded controlled study including infants at the age of 6 months was conducted. Infants were assigned randomly to either follow-on formula supplemented with L. fermentum plus galactooligosaccharide (experimental group, EG), or the same formula supplemented with only galactooligosaccharide (control group, CG). The main outcome was the incidence of infections for the 6-month duration of the study. RESULTS: The EG showed a significant 46% reduction in the incidence rate (IR) of gastrointestinal infections (EG: 0.196 ±â€Š0.51, CG: 0.363 ±â€Š0.53, IR ratio 0.54, 95% confidence interval [CI] 0.307-0.950, P = 0.032), 27% reduction in the incidence of upper respiratory tract infections (EG: 0.969 ±â€Š0.96, CG: 1.330 ±â€Š1.23, IR ratio 0.729, 95% CI 0.46-1.38, P = 0.026), and 30% reduction in the total number of infections (EG: 1.464 ±â€Š1.15, CG: 2.077 ±â€Š1.59, IR ratio 0.70, 95% CI 0.46-1.38, P = 0.003), at the end of the study period compared with CG. CONCLUSIONS: Administration of a follow-on formula with L. fermentum CECT5716 may be useful for the prevention of community-acquired gastrointestinal and upper respiratory infections.

Lactobacillus fermentum CECT 5716 is safe and well tolerated in infants of 1-6 months of age: a randomized controlled trial.

The objective of the study was to evaluate the safety and tolerance of an infant formula supplemented with Lactobacillus fermentum CECT5716, a probiotic strain isolated from breast milk, in infants of 1-6 months of age. A randomized double blinded controlled study including healthy infants was conducted. One month aged infants received a prebiotic infant formula supplemented with L. fermentum (experimental group) or the same formula without the probiotic strain (control group) for 5 months. The primary outcome of the study was average daily weight gain between baseline and 4 months of age. Secondary outcomes were other anthropometric data (length and head circumference), formula consumption, and tolerance. Incidence of infections was also recorded by pediatricians. No significant differences in weight gain were observed between both groups, neither at 4 months of age (29.0±7.8 vs 28.9±5.7g/day) nor at 6 months (25.1±6.1 vs 24.7±5.2g/day). There were no statistically significant differences in the consumption of the formulae or symptoms related to the tolerance of the formula. The incidence rate of gastrointestinal infections in infants of the control group was 3 times higher than in the probiotic group (p=0.018). Therefore, consumption of a prebiotic infant formula enriched with the human milk probiotic strain L. fermentum CECT5716 from 1 to 6 months of life is well tolerated and safe. Furthermore, the consumption of this formula may improve the health of the infants by reducing the incidence of gastrointestinal infections.

Intestinal and immunological effects of daily oral administration of Lactobacillus salivarius CECT5713 to healthy adults.

There is an increasing interest in the intestinal and immunological effects of probiotics. The aim of the present study is to evaluate the tolerance and beneficial effects in healthy adults of the strain, Lactobacillus salivarius CECT5713 isolated from breast milk. A phase II, randomized, double-blinded, placebo-controlled human clinical trial was carried out in 40 healthy adults. The Probiotic group received a daily dose of 2 x 10(8) CFU of L. salivarius CECT5713 in capsules during 4 weeks while volunteers of the control received only a placebo. Gastrointestinal and immunological parameters were analyzed. Results showed that L. salivarius CECT5713 was well tolerated and no adverse effects were detected. Consumption of the probiotic strain increased fecal lactobacilli counts (7.9+/-0.1 vs. 7.05+/-0.2 CFU/g feces, P=0.001). Also, an improvement in the frequency of defecation (P=0.04) was observed. Probiotic treatment induced significantly the percentage of NK cells and monocytes, as well as the plasmatic levels of immunoglobulins M, A and G, and the regulatory cytokine IL-10 (72.3+/-11.7 in probiotic group vs. 27.3+/-6.4 pg/mL in control group, P<0.01). Thus, it can be concluded that daily administration of L. salivarius CECT5713 to healthy adults is safe and improve gut microbiota and different parameters related to immune response.

Safety and tolerance of the human milk probiotic strain Lactobacillus salivarius CECT5713 in 6-month-old children.

OBJECTIVE: Intestinal microbiota plays an important role in the prevention of certain diseases during the pediatric years. Thus, there is an increasing interest in the addition of probiotics to infant formulas. The aim of this study was to evaluate the safety of a follow-on formula with Lactobacillus salivarius CECT5713 in 6-mo-old children. METHODS: The antibiotic susceptibility of L. salivarius CECT5713 was analyzed by a dilution method. A double-blinded, randomized, placebo controlled study was performed. Children (n = 80) were distributed in two groups and consumed the formula supplemented or not with probiotics (2 × 10(6) colony-forming units [cfu]/g) during 6 mo. Fecal samples were collected at enrollment, at 3 mo, and at the end of trial. Clinical and anthropometric evaluations were performed. Depending on the variable, one-way or two-way repeated measures analysis of variance were used for the statistical analysis. RESULTS: The antibiotic susceptibility profile of the strain resulted as safe. No adverse effects associated with the consumption of the probiotic formula were reported. In addition, clinical parameters did not differ between groups. Consumption of the probiotic supplemented formula led to an increase in the fecal lactobacilli content (7.6 ± 0.2 versus 7.9 ± 0.1 log cfu/g, P < 0.05). Lactobacillus salivarius CECT5713 was detected in the feces of volunteers from the probiotic group. Probiotic consumption induced a significant increase in the fecal concentration of butyric acid at 6 mo. CONCLUSION: Thus, a follow-on formula with L. salivarius CECT5713 is safe and well tolerated in 6-mo-old infants.

A probiotic dairy product containing L. gasseri CECT5714 and L. coryniformis CECT5711 induces immunological changes in children suffering from allergy.

The increase in the prevalence of allergic diseases in children has been attributed to an unbalanced immune response probably due to environmental factors. The immunoregulatory properties of probiotic bacteria could balance the disequilibrium in the immune response causing the allergic response. The aim of this study was to evaluate the immunological effects of the consumption of a dairy product containing two probiotic strains in children suffering from allergy. A double-blinded, randomized, control comparative study was performed with 44 allergic children. Children were randomly distributed in two groups, a control Yogurt and a Probiotic group. Both groups daily consumed 200 ml of a dairy fermented product for 3 months. The Yogurt group consumed a conventional yogurt, whereas the Probiotic group consumed a similar dairy product where Lactobacillus bulgaricus was substituted by a mixture of Lactobacillus gasseri CECT5714 and Lactobacillus coryniformis CECT5711 (at least 10(6) cfu/g each strain). Intestinal and immunological parameters were measured in fecal and blood samples. The consumption of the probiotic product induced a significant decrease in the level of IgE in plasma (p = 0.03) and an increase in CD4(+)/CD25(+) T regulatory cells (p = 0.01). The decrease in IgE was accompanied by a significant increase in mucosal IgA (p = 0.01). However, changes in other effector cells potentially involved in allergic reactions such as eosinophiles, basophiles or other IgE+ cells were not detected. The consumption of the probiotic product also induced significant changes in innate response as a significant increase in natural killer cells was detected (p = 0.03). The daily consumption of a probiotic product containing L. gasseri CECT5714 and L. coryniformis CECT5711 for 3 months induces, in allergic children, beneficial effects on immune parameters involved in the allergic response such as a reduction of IgE in plasma and an increase in regulatory T cells. The probiotic product also enhanced innate and specific immune parameters that may improve the general health status of children.

Safety assessment of Lactobacillus fermentum CECT5716, a probiotic strain isolated from human milk.

Lactobacillus fermentum CECT5716, a probiotic strain isolated from human milk, was characterized in a previous study. The objective of this study was to evaluate its sensitivity to antibiotics and its potential toxicity and translocation ability after oral administration to mice. For this purpose, 40 Balb/C mice were divided in two groups (n=20 per group). One group was treated orally with 10(10) colony forming units (cfu)/mouse/day of Lb. fermentum CECT5716 during 28 d. The other group only received the excipient and was used as control. Food intake, body weight, bacterial translocation and different biochemical and haematological parameters were analysed. Oral administration of Lb. fermentum CECT5716 to mice had no adverse effects on mice. There were no significant differences in body weight or food intake between control and probiotic-treated mice. No bacteraemia was observed and there was no treatment-associated bacterial translocation to liver or spleen. Stress oxidative markers were not different in control and probiotic-treated mice. These results suggest that the strain Lb. fermentum CECT5716 is non-pathogenic for mice even in doses 10,000 times higher (expressed per kg of body weight) than those normally consumed by humans.

Dietary eicosapentaenoic acid and docosahexaenoic acid equally incorporate as decosahexaenoic acid but differ in inflammatory effects.

OBJECTIVE: The omega-3 polyunsaturated fatty acids are involved in the modulation of the immune response. Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) produced from dietary precursors may not be sufficient to match nutritional requirements and thus should be included in our diet. In this sense, the administration of higher amounts of DHA than of EPA in infant formulations is recommended. The aims of this work were to demonstrate that dietary administration of EPA or DHA to mice allows reaching similar tissue DHA levels and to compare their anti-inflammatory effects and mechanisms of action. METHODS: Balb/c mice were fed diets enriched with EPA or DHA for 3 wk. Twelve hours before sacrifice, a contact dermatitis was induced in the ears of the animals. Tissue fatty acid contents were determined. Cytokine and immunoglobulin concentrations were measured by enzyme-linked immunosorbent assay, and ears were collected to analyze local inflammatory effects. RESULTS: The DHA concentrations attained in tissues were similar to the two diets, whereas the EPA concentration increased only when the diet was enriched with this polyunsaturated fatty acid. Although EPA and DHA reduced ear inflammation, EPA reduced neutrophil infiltration in the ears more efficiently. EPA was associated with a greater reduction in the systemic macrophage inflammatory response and T-helper type 2 response and with increased interleukin-10 production. CONCLUSION: Similar levels of DHA in tissues are reached in mice fed an EPA- or a DHA-enriched diet. Dietary EPA and DHA show anti-inflammatory properties, but EPA appears to be more potent.

Beneficial effects of probiotic bacteria isolated from breast milk.

Breast milk is the best food for the neonate because it provides a unique combination of proteins, carbohydrates, lipids, minerals and vitamins that ensures the correct growth and development of the infant. In addition, it also contains bioactive compounds responsible for a wide range of beneficial effects such as the promotion of immune system maturation and the protection against infections. Among these bioactive agents, probiotic bacteria have been recently isolated from human milk. The present work reviews the beneficial effects of these bacteria both in animal models and in clinical trials. The promotion of immune system maturation and defence against infections as well as the anti-inflammatory properties are among the main healthy effects of these bacteria. The isolation of probiotic bacteria with beneficial effects for the host provides scientific support for the supplementation of infant formula with these bacteria, in order to advance the pursuit of the main goal of formula: to mimic breast milk and its functional effects as closely as possible.

Oral intake of Lactobacillus fermentum CECT5716 enhances the effects of influenza vaccination.

OBJECTIVE: We studied the coadjuvant capability of oral consumption of the breast-milk-isolated strain Lactobacillus fermentum (CECT5716) for an anti-influenza vaccine. METHODS: A randomized, double-blinded, placebo-controlled human clinical trial including 50 volunteers (31 male and 19 female) was performed to address the immunologic effects of an intramuscular anti-influenza vaccine in adults (33.0 +/- 7.7 y old). Fifty percent of volunteers received an oral daily dose of methylcellulose (placebo) or probiotic bacteria (1 x 10(10) colony-forming units/d) 2 wk before vaccination and 2 wk after vaccination. RESULTS: Two weeks after vaccination there was an increase in the proportion of natural killer cells in the probiotic group but not in the placebo group. The vaccination induced an increase in T-helper type 1 cytokine concentrations and in T-helper and T-cytotoxic proportions in both groups; however, the probiotic group showed a significant higher induction in some of these parameters. Regarding the humoral effects, induction of antibody response in the placebo group could not be detected. In the case of the probiotic group, a significant increase in antigen specific immunoglobulin A was detected. Although an increase in total immunoglobulin M was observed, changes in anti-influenza antigen specific immunoglobulin M were not observed. The incidence of an influenza-like illness during 5 mo after vaccination (October to February) was lower in the group consuming the probiotic bacteria. CONCLUSION: Oral administration of the strain L. fermentum CECT5716 potentates the immunologic response of an anti-influenza vaccine and may provide enhanced systemic protection from infection by increasing the T-helper type 1 response and virus-neutralizing antibodies.

A comparative study of the preventative effects exerted by two probiotics, Lactobacillus reuteri and Lactobacillus fermentum, in the trinitrobenzenesulfonic acid model of rat colitis.

The intestinal anti-inflammatory effects of two probiotics isolated from breast milk, Lactobacillus reuteri and L. fermentum, were evaluated and compared in the trinitrobenzenesulfonic acid (TNBS) model of rat colitis. Colitis was induced in rats by intracolonic administration of 10 mg TNBS dissolved in 50% ethanol (0.25 ml). Either L. reuteri or L. fermentum was daily administered orally (5 x 10(8) colony-forming units suspended in 0.5 ml skimmed milk) to each group of rats (n 10) for 3 weeks, starting 2 weeks before colitis induction. Colonic damage was evaluated histologically and biochemically, and the colonic luminal contents were used for bacterial studies and for SCFA production. Both probiotics showed intestinal anti-inflammatory effects in this model of experimental colitis, as evidenced histologically and by a significant reduction of colonic myeloperoxidase activity (P<0.05). L. fermentum significantly counteracted the colonic glutathione depletion induced by the inflammatory process. In addition, both probiotics lowered colonic TNFalpha levels (P<0.01) and inducible NO synthase expression when compared with non-treated rats; however, the decrease in colonic cyclo-oxygenase-2 expression was only achieved with L.fermentum administration. Finally, the two probiotics induced the growth of Lactobacilli species in comparison with control colitic rats, but the production of SCFA in colonic contents was only increased when L. fermentum was given. In conclusion, L. fermentum can exert beneficial immunomodulatory properties in inflammatory bowel disease, being more effective than L. reuteri, a probiotic with reputed efficacy in promoting beneficial effects on human health.